#METABOLOMICS WORKBENCH Sethjparker_20250619_131136 DATATRACK_ID:6063 STUDY_ID:ST004016 ANALYSIS_ID:AN006621 PROJECT_ID:PR002501
VERSION             	1
CREATED_ON             	July 2, 2025, 11:15 am
#PROJECT
PR:PROJECT_TITLE                 	Metabolomics analysis of SNAT2-deficient cells: implications for the discovery
PR:PROJECT_TITLE                 	of selective transporter inhibitors
PR:PROJECT_TYPE                  	Manuscript
PR:PROJECT_SUMMARY               	Amino acid uptake by the solute carrier family of transporter proteins is
PR:PROJECT_SUMMARY               	critical to support cell metabolism, and inhibition of transporter activity
PR:PROJECT_SUMMARY               	represents a tractable strategy to restrict nutrient availability to cancer
PR:PROJECT_SUMMARY               	cells. A small molecule inhibitor of the sodium-coupled neutral amino acid
PR:PROJECT_SUMMARY               	transporter 2 (SNAT2),
PR:PROJECT_SUMMARY               	3-(N-methyl(4-methylphenyl)sulfonamido)-N-(2-trifluoromethylbenzyl)thiophene-2-carboxamide
PR:PROJECT_SUMMARY               	(MMTC/57E), was recently identified and was shown to inhibit cell proliferation
PR:PROJECT_SUMMARY               	when combined with glucose transport inhibitors in breast and pancreatic cancer
PR:PROJECT_SUMMARY               	cell lines. In this study, we use mass spectrometry-based metabolomics and
PR:PROJECT_SUMMARY               	establish cell-based assays for the SNAT2 transporter. We show that SNAT2
PR:PROJECT_SUMMARY               	knockout cells have significant defects in amino acid availability. Using our
PR:PROJECT_SUMMARY               	established assays, we fail to observe that MMTC/57E inhibits SNAT2 activity
PR:PROJECT_SUMMARY               	likely due to its poor solubility.
PR:INSTITUTE                     	University of British Columbia
PR:DEPARTMENT                    	Biochemistry & Molecular Biology
PR:LABORATORY                    	Parker laboratory
PR:LAST_NAME                     	Parker
PR:FIRST_NAME                    	Seth
PR:ADDRESS                       	950 W 28th Ave, Vancouver, British Columbia, V6H 0B3, Canada
PR:EMAIL                         	seth.parker@bcchr.ca
PR:PHONE                         	6048753121
#STUDY
ST:STUDY_TITLE                   	Evaluation of SNAT2-dependent and sodium-dependent MeAIB uptake using GCMS
ST:STUDY_SUMMARY                 	HY15549 cell lines expressing a dox-inducible SNAT2 cDNA and lacking endogenous
ST:STUDY_SUMMARY                 	Snat2 expression were plated and evaluated for MeAIB uptake by culturing cells
ST:STUDY_SUMMARY                 	with a gradient from 0.125-4 mM for 30-minutes in PBS for 30-minutes prior to
ST:STUDY_SUMMARY                 	GCMS analysis. For sodium-deficient uptake experiments, PBS was prepared using
ST:STUDY_SUMMARY                 	equimolar concentrations of N-methyl-D-glucamine to replace sodium chloride;
ST:STUDY_SUMMARY                 	potassium salts were used to replace other sodium salts. SNAT2 exhibited
ST:STUDY_SUMMARY                 	saturable uptake kinetics with an apparent Km of ~ were engineered to stably
ST:STUDY_SUMMARY                 	express a human SNAT1 cDNA or an empty vector (EV, controls) by lentiviral
ST:STUDY_SUMMARY                 	transduction. Either EV or +SNAT1 cell lines were plated and evaluated for MeAIB
ST:STUDY_SUMMARY                 	uptake by culturing cells with a gradient from 0.125-4 mM for 30-minutes in PBS
ST:STUDY_SUMMARY                 	for 30-minutes prior to GCMS analysis. SNAT1 exhibited saturable uptake kinetics
ST:STUDY_SUMMARY                 	with an apparent Km of ~0.34 mM and was significantly sodium dependent.
ST:INSTITUTE                     	University of British Columbia
ST:DEPARTMENT                    	Biochemistry & Molecular Biology
ST:LABORATORY                    	Parker laboratory
ST:LAST_NAME                     	Parker
ST:FIRST_NAME                    	Seth
ST:ADDRESS                       	950 W 28th Ave, Vancouver, British Columbia, V6H 0B3, Canada
ST:EMAIL                         	seth.parker@bcchr.ca
ST:PHONE                         	6048753121
#SUBJECT
SU:SUBJECT_TYPE                  	Mammal
SU:SUBJECT_SPECIES               	Mus musculus
SU:TAXONOMY_ID                   	10090
SU:GENDER                        	Female
#SUBJECT_SAMPLE_FACTORS:         	SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data
SUBJECT_SAMPLE_FACTORS           	1	HYiSNAT2_noDox_wNa_M125_3	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.125	Replicate=3; RAW_FILE_NAME(Raw file name)=HYiSNAT2_noDox_wNa_M125_3.CDF
SUBJECT_SAMPLE_FACTORS           	2	HYiSNAT2_noDox_wNa_M125_r1	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.125	Replicate=1; RAW_FILE_NAME(Raw file name)=HYiSNAT2_noDox_wNa_M125_r1.CDF
SUBJECT_SAMPLE_FACTORS           	3	HYiSNAT2_noDox_wNa_M125_r2	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.125	Replicate=2; RAW_FILE_NAME(Raw file name)=HYiSNAT2_noDox_wNa_M125_r2.CDF
SUBJECT_SAMPLE_FACTORS           	4	HYiSNAT2_noDox_wNa_M250_3	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.25	Replicate=3; RAW_FILE_NAME(Raw file name)=HYiSNAT2_noDox_wNa_M250_3.CDF
SUBJECT_SAMPLE_FACTORS           	5	HYiSNAT2_noDox_wNa_M250_r1	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.25	Replicate=1; RAW_FILE_NAME(Raw file name)=HYiSNAT2_noDox_wNa_M250_r1.CDF
SUBJECT_SAMPLE_FACTORS           	6	HYiSNAT2_noDox_wNa_M250_r2	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.25	Replicate=2; RAW_FILE_NAME(Raw file name)=HYiSNAT2_noDox_wNa_M250_r2.CDF
SUBJECT_SAMPLE_FACTORS           	7	HYiSNAT2_noDox_wNa_M500_3	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.5	Replicate=3; RAW_FILE_NAME(Raw file name)=HYiSNAT2_noDox_wNa_M500_3.CDF
SUBJECT_SAMPLE_FACTORS           	8	HYiSNAT2_noDox_wNa_M500_r1	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.5	Replicate=1; RAW_FILE_NAME(Raw file name)=HYiSNAT2_noDox_wNa_M500_r1.CDF
SUBJECT_SAMPLE_FACTORS           	9	HYiSNAT2_noDox_wNa_M500_r2	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.5	Replicate=2; RAW_FILE_NAME(Raw file name)=HYiSNAT2_noDox_wNa_M500_r2.CDF
SUBJECT_SAMPLE_FACTORS           	10	HYiSNAT2_noDox_wNa_M1000_3	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):1	Replicate=3; RAW_FILE_NAME(Raw file name)=HYiSNAT2_noDox_wNa_M1000_3.CDF
SUBJECT_SAMPLE_FACTORS           	11	HYiSNAT2_noDox_wNa_M1000_r1	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):1	Replicate=1; RAW_FILE_NAME(Raw file name)=HYiSNAT2_noDox_wNa_M1000_r1.CDF
SUBJECT_SAMPLE_FACTORS           	12	HYiSNAT2_noDox_wNa_M1000_r2	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):1	Replicate=2; RAW_FILE_NAME(Raw file name)=HYiSNAT2_noDox_wNa_M1000_r2.CDF
SUBJECT_SAMPLE_FACTORS           	13	HYiSNAT2_noDox_wNa_M2000_3	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):2	Replicate=3; RAW_FILE_NAME(Raw file name)=HYiSNAT2_noDox_wNa_M2000_3.CDF
SUBJECT_SAMPLE_FACTORS           	14	HYiSNAT2_noDox_wNa_M2000_r1	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):2	Replicate=1; RAW_FILE_NAME(Raw file name)=HYiSNAT2_noDox_wNa_M2000_r1.CDF
SUBJECT_SAMPLE_FACTORS           	15	HYiSNAT2_noDox_wNa_M2000_r2	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):2	Replicate=2; RAW_FILE_NAME(Raw file name)=HYiSNAT2_noDox_wNa_M2000_r2.CDF
SUBJECT_SAMPLE_FACTORS           	16	HYiSNAT2_noDox_wNa_M4000_3	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):4	Replicate=3; RAW_FILE_NAME(Raw file name)=HYiSNAT2_noDox_wNa_M4000_3.CDF
SUBJECT_SAMPLE_FACTORS           	17	HYiSNAT2_noDox_wNa_M4000_r1	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):4	Replicate=1; RAW_FILE_NAME(Raw file name)=HYiSNAT2_noDox_wNa_M4000_r1.CDF
SUBJECT_SAMPLE_FACTORS           	18	HYiSNAT2_noDox_wNa_M4000_r2	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):4	Replicate=2; RAW_FILE_NAME(Raw file name)=HYiSNAT2_noDox_wNa_M4000_r2.CDF
SUBJECT_SAMPLE_FACTORS           	19	HYiSNAT2_wDox_noNa_M125_3	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.125	Replicate=3; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_noNa_M125_3.CDF
SUBJECT_SAMPLE_FACTORS           	20	HYiSNAT2_wDox_noNa_M125_r1	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.125	Replicate=1; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_noNa_M125_r1.CDF
SUBJECT_SAMPLE_FACTORS           	21	HYiSNAT2_wDox_noNa_M125_r2	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.125	Replicate=2; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_noNa_M125_r2.CDF
SUBJECT_SAMPLE_FACTORS           	22	HYiSNAT2_wDox_noNa_M250_3	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.25	Replicate=3; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_noNa_M250_3.CDF
SUBJECT_SAMPLE_FACTORS           	23	HYiSNAT2_wDox_noNa_M250_r1	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.25	Replicate=1; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_noNa_M250_r1.CDF
SUBJECT_SAMPLE_FACTORS           	24	HYiSNAT2_wDox_noNa_M250_r2	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.25	Replicate=2; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_noNa_M250_r2.CDF
SUBJECT_SAMPLE_FACTORS           	25	HYiSNAT2_wDox_noNa_M500_3	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.5	Replicate=3; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_noNa_M500_3.CDF
SUBJECT_SAMPLE_FACTORS           	26	HYiSNAT2_wDox_noNa_M500_r1	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.5	Replicate=1; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_noNa_M500_r1.CDF
SUBJECT_SAMPLE_FACTORS           	27	HYiSNAT2_wDox_noNa_M500_r2	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.5	Replicate=2; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_noNa_M500_r2.CDF
SUBJECT_SAMPLE_FACTORS           	28	HYiSNAT2_wDox_noNa_M1000_3	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):1	Replicate=3; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_noNa_M1000_3.CDF
SUBJECT_SAMPLE_FACTORS           	29	HYiSNAT2_wDox_noNa_M1000_r1	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):1	Replicate=1; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_noNa_M1000_r1.CDF
SUBJECT_SAMPLE_FACTORS           	30	HYiSNAT2_wDox_noNa_M1000_r2	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):1	Replicate=2; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_noNa_M1000_r2.CDF
SUBJECT_SAMPLE_FACTORS           	31	HYiSNAT2_wDox_noNa_M2000_3	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):2	Replicate=3; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_noNa_M2000_3.CDF
SUBJECT_SAMPLE_FACTORS           	32	HYiSNAT2_wDox_noNa_M2000_r1	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):2	Replicate=1; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_noNa_M2000_r1.CDF
SUBJECT_SAMPLE_FACTORS           	33	HYiSNAT2_wDox_noNa_M2000_r2	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):2	Replicate=2; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_noNa_M2000_r2.CDF
SUBJECT_SAMPLE_FACTORS           	34	HYiSNAT2_wDox_noNa_M4000_3	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):4	Replicate=3; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_noNa_M4000_3.CDF
SUBJECT_SAMPLE_FACTORS           	35	HYiSNAT2_wDox_noNa_M4000_r1	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):4	Replicate=1; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_noNa_M4000_r1.CDF
SUBJECT_SAMPLE_FACTORS           	36	HYiSNAT2_wDox_noNa_M4000_r2	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):4	Replicate=2; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_noNa_M4000_r2.CDF
SUBJECT_SAMPLE_FACTORS           	37	HYiSNAT2_wDox_wNa_M125_3	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.125	Replicate=3; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_wNa_M125_3.CDF
SUBJECT_SAMPLE_FACTORS           	38	HYiSNAT2_wDox_wNa_M125_r1	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.125	Replicate=1; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_wNa_M125_r1.CDF
SUBJECT_SAMPLE_FACTORS           	39	HYiSNAT2_wDox_wNa_M125_r2	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.125	Replicate=2; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_wNa_M125_r2.CDF
SUBJECT_SAMPLE_FACTORS           	40	HYiSNAT2_wDox_wNa_M250_3	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.25	Replicate=3; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_wNa_M250_3.CDF
SUBJECT_SAMPLE_FACTORS           	41	HYiSNAT2_wDox_wNa_M250_r1	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.25	Replicate=1; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_wNa_M250_r1.CDF
SUBJECT_SAMPLE_FACTORS           	42	HYiSNAT2_wDox_wNa_M250_r2	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.25	Replicate=2; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_wNa_M250_r2.CDF
SUBJECT_SAMPLE_FACTORS           	43	HYiSNAT2_wDox_wNa_M500_3	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.5	Replicate=3; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_wNa_M500_3.CDF
SUBJECT_SAMPLE_FACTORS           	44	HYiSNAT2_wDox_wNa_M500_r1	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.5	Replicate=1; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_wNa_M500_r1.CDF
SUBJECT_SAMPLE_FACTORS           	45	HYiSNAT2_wDox_wNa_M500_r2	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.5	Replicate=2; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_wNa_M500_r2.CDF
SUBJECT_SAMPLE_FACTORS           	46	HYiSNAT2_wDox_wNa_M1000_3	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):1	Replicate=3; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_wNa_M1000_3.CDF
SUBJECT_SAMPLE_FACTORS           	47	HYiSNAT2_wDox_wNa_M1000_r1	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):1	Replicate=1; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_wNa_M1000_r1.CDF
SUBJECT_SAMPLE_FACTORS           	48	HYiSNAT2_wDox_wNa_M1000_r2	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):1	Replicate=2; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_wNa_M1000_r2.CDF
SUBJECT_SAMPLE_FACTORS           	49	HYiSNAT2_wDox_wNa_M2000_3	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):2	Replicate=3; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_wNa_M2000_3.CDF
SUBJECT_SAMPLE_FACTORS           	50	HYiSNAT2_wDox_wNa_M2000_r1	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):2	Replicate=1; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_wNa_M2000_r1.CDF
SUBJECT_SAMPLE_FACTORS           	51	HYiSNAT2_wDox_wNa_M2000_r2	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):2	Replicate=2; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_wNa_M2000_r2.CDF
SUBJECT_SAMPLE_FACTORS           	52	HYiSNAT2_wDox_wNa_M4000_3	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):4	Replicate=3; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_wNa_M4000_3.CDF
SUBJECT_SAMPLE_FACTORS           	53	HYiSNAT2_wDox_wNa_M4000_r1	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):4	Replicate=1; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_wNa_M4000_r1.CDF
SUBJECT_SAMPLE_FACTORS           	54	HYiSNAT2_wDox_wNa_M4000_r2	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):4	Replicate=2; RAW_FILE_NAME(Raw file name)=HYiSNAT2_wDox_wNa_M4000_r2.CDF
#COLLECTION
CO:COLLECTION_SUMMARY            	Cells were washed twice with room temperature (20°C) PBS before adding 2 mL of
CO:COLLECTION_SUMMARY            	PBS containing 0.125-4 mM MeAIB. After a 30-minute incubation at 20°C, the
CO:COLLECTION_SUMMARY            	transport buffer was aspirated and the cells were washed three times with ice
CO:COLLECTION_SUMMARY            	cold 0.9% NaCl. To extract metabolites, 1 mL of ice cold 80% methanol containing
CO:COLLECTION_SUMMARY            	2.5 nmol of 13C3-alanine was added to each well. The plates were shaken at ~500
CO:COLLECTION_SUMMARY            	rpm for 15 minutes at room temperature and 900 µL was transferred to a new tube
CO:COLLECTION_SUMMARY            	and dried using a SpeedVac for GCMS analysis.
CO:SAMPLE_TYPE                   	Tumor cells
#TREATMENT
TR:TREATMENT_SUMMARY             	No treatment.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Dried samples were derivatized for GCMS analysis. 25 µL of methoxyamine
SP:SAMPLEPREP_SUMMARY            	hydrochloride (20 mg/mL, prepared in pyridine) was added to each tube, which was
SP:SAMPLEPREP_SUMMARY            	vortexed and incubated at 37°C for 60 minutes. Following the incubation, 25 µL
SP:SAMPLEPREP_SUMMARY            	of MTBSTFA + 1% TBDMSCl was added to each tube, which was vortexed and
SP:SAMPLEPREP_SUMMARY            	centrifuged at 21,300 x g for 15-minutes at 20°C. 40 µL of the resulting
SP:SAMPLEPREP_SUMMARY            	supernatant was transferred to a plastic sample vial for GCMS analysis.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	Agilent DB35-MS (30m x 0.25mm, 0.25µm) was interfaced with an inert column (5m x
CH:CHROMATOGRAPHY_SUMMARY        	0.15mm, 0 µm) to the MSD. Initial oven temperature maintained at 75°C for 3
CH:CHROMATOGRAPHY_SUMMARY        	minutes, ramped to 255°C at 7.5°C/min, held for 10 minutes, ramped to 320°C at 10
CH:CHROMATOGRAPHY_SUMMARY        	C/min, held for 5 minutes. The inlet was maintained at 270°C in splitless mode;
CH:CHROMATOGRAPHY_SUMMARY        	an injection volume of 1 uL was used for all samples.
CH:CHROMATOGRAPHY_TYPE           	GC
CH:INSTRUMENT_NAME               	Agilent 8890
CH:COLUMN_NAME                   	Agilent DB-35MS (30m x 0.25mm, 0.25µm) interfaced with an inert column (5m x 0.15mm, 0 µm)
CH:SOLVENT_A                     	UHP Helium
CH:SOLVENT_B                     	None
CH:FLOW_GRADIENT                 	100% Helium
CH:FLOW_RATE                     	1 mL/min
CH:COLUMN_TEMPERATURE            	75°C
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Agilent 5977B
MS:INSTRUMENT_TYPE               	Single quadrupole
MS:MS_TYPE                       	EI
MS:ION_MODE                      	POSITIVE
MS:MS_COMMENTS                   	The MS was operated in full-scan mode from 100 to 650 m/z with a threshold of
MS:MS_COMMENTS                   	150, a scan speed of 781 u/sec, a frequency of 1.4 scans/sec, a cycle time of
MS:MS_COMMENTS                   	723 ms, and a step size of 0.1 m/z. The MS Source and MS Quad temperatures were
MS:MS_COMMENTS                   	set to 230°C and 150°C, respectively. A gain factor of 1 was used.
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	ion counts
MS_METABOLITE_DATA_START
Samples	HYiSNAT2_noDox_wNa_M125_r1	HYiSNAT2_noDox_wNa_M250_r1	HYiSNAT2_noDox_wNa_M500_r1	HYiSNAT2_noDox_wNa_M1000_r1	HYiSNAT2_noDox_wNa_M2000_r1	HYiSNAT2_noDox_wNa_M4000_r1	HYiSNAT2_wDox_noNa_M125_r1	HYiSNAT2_wDox_noNa_M250_r1	HYiSNAT2_wDox_noNa_M500_r1	HYiSNAT2_wDox_noNa_M1000_r1	HYiSNAT2_wDox_noNa_M2000_r1	HYiSNAT2_wDox_noNa_M4000_r1	HYiSNAT2_wDox_wNa_M125_r1	HYiSNAT2_wDox_wNa_M250_r1	HYiSNAT2_wDox_wNa_M500_r1	HYiSNAT2_wDox_wNa_M1000_r1	HYiSNAT2_wDox_wNa_M2000_r1	HYiSNAT2_wDox_wNa_M4000_r1	HYiSNAT2_noDox_wNa_M125_r2	HYiSNAT2_noDox_wNa_M250_r2	HYiSNAT2_noDox_wNa_M500_r2	HYiSNAT2_noDox_wNa_M1000_r2	HYiSNAT2_noDox_wNa_M2000_r2	HYiSNAT2_noDox_wNa_M4000_r2	HYiSNAT2_wDox_noNa_M125_r2	HYiSNAT2_wDox_noNa_M250_r2	HYiSNAT2_wDox_noNa_M500_r2	HYiSNAT2_wDox_noNa_M1000_r2	HYiSNAT2_wDox_noNa_M2000_r2	HYiSNAT2_wDox_noNa_M4000_r2	HYiSNAT2_wDox_wNa_M125_r2	HYiSNAT2_wDox_wNa_M250_r2	HYiSNAT2_wDox_wNa_M500_r2	HYiSNAT2_wDox_wNa_M1000_r2	HYiSNAT2_wDox_wNa_M2000_r2	HYiSNAT2_wDox_wNa_M4000_r2	HYiSNAT2_noDox_wNa_M125_3	HYiSNAT2_noDox_wNa_M250_3	HYiSNAT2_noDox_wNa_M500_3	HYiSNAT2_noDox_wNa_M1000_3	HYiSNAT2_noDox_wNa_M2000_3	HYiSNAT2_noDox_wNa_M4000_3	HYiSNAT2_wDox_noNa_M125_3	HYiSNAT2_wDox_noNa_M250_3	HYiSNAT2_wDox_noNa_M500_3	HYiSNAT2_wDox_noNa_M1000_3	HYiSNAT2_wDox_noNa_M2000_3	HYiSNAT2_wDox_noNa_M4000_3	HYiSNAT2_wDox_wNa_M125_3	HYiSNAT2_wDox_wNa_M250_3	HYiSNAT2_wDox_wNa_M500_3	HYiSNAT2_wDox_wNa_M1000_3	HYiSNAT2_wDox_wNa_M2000_3	HYiSNAT2_wDox_wNa_M4000_3
Factors	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.125	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.25	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.5	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):1	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):2	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):4	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.125	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.25	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.5	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):1	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):2	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):4	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.125	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.25	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.5	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):1	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):2	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):4	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.125	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.25	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.5	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):1	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):2	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):4	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.125	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.25	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.5	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):1	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):2	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):4	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.125	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.25	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.5	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):1	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):2	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):4	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.125	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.25	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.5	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):1	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):2	Factor:with Sodium | Doxycycline (y/n):no | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):4	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.125	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.25	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.5	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):1	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):2	Factor:no Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):4	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.125	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.25	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):0.5	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):1	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):2	Factor:with Sodium | Doxycycline (y/n):yes | Sample source:HY15549 Pancreatic Cancer Cells | MeAIB concentration (mM):4
MeAIB_174	405332	387482	524669	954897	947018	1305911	154935	116618	235247	369823	695904	958723	4197317	5631749	9462925	11222144	12179808	12492380	315566	395520	503022	573824	868522	1626845	166488	223907	305551	474063	800439	1609009	4197452	6113548	8019159	13349665	11239654	12083929	181066	301013	394727	574177	786183	1325073	93180	141202	220872	448323	988863	2023769	3329687	7562631	8143597	13602522	13578978	16935590
Ala_232	7680209	5255180	6979055	7903397	4340534	5757151	8872208	6116230	7572446	6927811	9071961	8118506	17572921	17919860	22691432	24475664	20792635	24498165	4341681	3420983	5439138	5101192	5289860	3933773	8539400	7998969	11701740	10849862	8980797	7105906	14997204	12664449	20027591	27174744	28053892	25439982	4124761	3654114	6457043	6565665	6331732	5801647	10131865	8515541	11865028	11314657	12009533	12355423	13443880	16164301	20730089	26942787	31620180	36538063
Ala_260	6841458	4965108	6423890	7014196	4108817	5279181	8216407	5859956	7365670	6560928	8560370	7568509	16706743	17169063	22193208	24002641	20924477	24933780	3951121	3186072	4893938	4732805	4727499	3675530	7905822	7300057	10824175	10187433	8325188	6893553	14068349	12155008	19183824	26616629	27854719	25504440	3624698	3203773	5614792	5830622	5568052	5094021	9637204	8162295	11087484	10238589	10939672	11483385	12649127	15140568	19644683	25731045	30756148	35709572
Ala_264	3038291.498	2214438.168	2910664.559	3234245.776	2417627.923	2968483.476	744406.4742	572517.7012	685007.31	637066.1088	840628.334	668299.3447	2696468.32	2637168.077	2623237.186	2438668.326	1948068.809	2044569.96	2496713.36	1992888.036	2718093.165	2555714.7	2843590.649	2256040.314	667251.3768	639484.9932	887582.35	795638.5173	705143.4236	582505.2285	2409908.184	1862147.226	2014301.52	2512609.778	2281301.486	2099015.412	2398462.667	2198749.41	2842207.71	2973617.22	3142051.744	2904101.372	792178.1688	721546.878	915826.1784	955260.3537	913462.612	927857.508	2061807.701	2269571.143	2443798.565	2745502.502	2574289.588	2871049.589
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	m/z	rt
MeAIB_174	174	13.64
Ala_232	232	18.18
Ala_260	260	18.18
Ala_264	264	18.18
METABOLITES_END
#END