#METABOLOMICS WORKBENCH araskind_20170802_132817 DATATRACK_ID:1182 STUDY_ID:ST000833 ANALYSIS_ID:AN001842 PROJECT_ID:PR000594 VERSION 1 CREATED_ON January 16, 2019, 9:11 am #PROJECT PR:PROJECT_TITLE Influence of murine norovirus on cell metabolism PR:PROJECT_TYPE MS analysis PR:PROJECT_SUMMARY C6 effects in RAW PR:INSTITUTE University of Michigan PR:DEPARTMENT Microbiology & Immunology PR:LABORATORY Passalacqua Lab PR:LAST_NAME Passalacqua PR:FIRST_NAME Karla PR:ADDRESS Ann Arbor, MI PR:EMAIL kpassal@umich.edu PR:PHONE 734-763-5639 #STUDY ST:STUDY_TITLE Influence of murine norovirus on cell metabolism ST:STUDY_TYPE MS analysis ST:STUDY_SUMMARY 6.5 X 10^6 cells were plated in 10 cm Tissue Culture dishes and allowed to ST:STUDY_SUMMARY recover overnight at 37 degrees and 5% CO2. In the am, supernatant was removed, ST:STUDY_SUMMARY and 12 mL of medium containing either Murine Norovirus-1 (MNV) at an MOI=5 or ST:STUDY_SUMMARY medium containing a v/v match of mock lysate was added to the cells. Plates were ST:STUDY_SUMMARY rocked on ice for 1 hour, then cells were washed 3X with cold DPBS++, and plain ST:STUDY_SUMMARY medium as added to cells. Plates were then incubated for 7.5 hours at 37 degrees ST:STUDY_SUMMARY and 5% CO2. Cells were washed with 12 mL of 150 mM Ammonium Acetate, swirled 8 ST:STUDY_SUMMARY times, and immediately quenched with liquid nitrogen. Cells were then frozen at ST:STUDY_SUMMARY -80 degrees. ST:INSTITUTE University of Michigan ST:DEPARTMENT Biomedical Research Core Facilities ST:LABORATORY Metabolomics core ST:LAST_NAME Kachman ST:FIRST_NAME Maureen ST:ADDRESS 6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714 ST:EMAIL mkachman@med.umich.edu ST:PHONE (734) 232-8175 ST:NUM_GROUPS 2 ST:TOTAL_SUBJECTS 1 ST:STUDY_COMMENTS BIOHAZARD - MURINE NOROVIRUS - SHOULD BE ENTIRELY NON-INFECTIOUS DUE TO THE ST:STUDY_COMMENTS LIQUID NITROGEN, BUT PLEASE HANDLE AND DISPOSE OF IN BIOHAZARD WASTE. IMPORTANT ST:STUDY_COMMENTS NOTE: I request that the metabolites of the TCA+ assay that are to be assessed ST:STUDY_COMMENTS quantitatively please be sure to include ATP, ADP, AMP, NAD, NADH, NADP, NADPH, ST:STUDY_COMMENTS E4P, S7P, 6PG, G3P H6P and Lactate. Also, I would request that both reduced and ST:STUDY_COMMENTS oxidized Glutathione as well as Fructose 1,6-bisphosphate be included in the ST:STUDY_COMMENTS non-quantitative data. #SUBJECT SU:SUBJECT_TYPE MOUSE SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS SU0015382 S00023719 Medium:mock lysate SUBJECT_SAMPLE_FACTORS SU0015382 S00023720 Medium:mock lysate SUBJECT_SAMPLE_FACTORS SU0015382 S00023721 Medium:mock lysate SUBJECT_SAMPLE_FACTORS SU0015382 S00023722 Medium:MNV infected SUBJECT_SAMPLE_FACTORS SU0015382 S00023723 Medium:MNV infected SUBJECT_SAMPLE_FACTORS SU0015382 S00023724 Medium:MNV infected #COLLECTION CO:COLLECTION_SUMMARY - CO:SAMPLE_TYPE Cultured cells #TREATMENT TR:TREATMENT_SUMMARY - #SAMPLEPREP SP:SAMPLEPREP_SUMMARY - SP:SAMPLEPREP_PROTOCOL_FILENAME A011_Glycolysis-TCA-nucleotides.pdf #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE GC CH:INSTRUMENT_NAME Agilent GC_7890N CH:COLUMN_NAME Agilent HP5-MS (15m × 0.25mm, 0.25 um) #ANALYSIS AN:ANALYSIS_TYPE MS AN:ANALYSIS_PROTOCOL_FILE A011_Glycolysis-TCA-nucleotides.pdf AN:ACQUISITION_PARAMETERS_FILE HEIDIFAMES.M #MS MS:INSTRUMENT_NAME Agilent MS_5975 MS:INSTRUMENT_TYPE Single quadrupole MS:MS_TYPE EI MS:ION_MODE POSITIVE MS:MS_COMMENTS none #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS pmol/ug protein MS_METABOLITE_DATA_START Samples S00023719 S00023720 S00023721 S00023722 S00023723 S00023724 Factors Medium:mock lysate Medium:mock lysate Medium:mock lysate Medium:MNV infected Medium:MNV infected Medium:MNV infected PYRUVIC ACID 9.301541105 14.90496856 14.99708502 11.80735452 9.116197265 5.77155209 FUMARIC ACID 36.17005854 38.38719496 26.84813761 53.70050442 33.11044567 19.60659917 LACTIC ACID 2435.484483 2288.154623 1724.146814 2678.46717 2016.551207 1280.271163 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name moverz_quant ri ri_type pubchem_id inchi_key kegg_id other_id other_id_type PYRUVIC ACID 1060 FUMARIC ACID 444972 LACTIC ACID 612 METABOLITES_END #END