#METABOLOMICS WORKBENCH FareseWalthersLab2022_20220917_145916 DATATRACK_ID:3464 STUDY_ID:ST002303 ANALYSIS_ID:AN003763 PROJECT_ID:PR001476 VERSION 1 CREATED_ON October 6, 2022, 9:40 am #PROJECT PR:PROJECT_TITLE Fitm2 is required for ER homeostasis and normal function of murine liver PR:PROJECT_SUMMARY The ER-resident protein fat-inducing transcript 2 (FIT2) catalyzes acyl-CoA PR:PROJECT_SUMMARY cleavage in vitro and is required for endoplasmic reticulum (ER) homeostasis and PR:PROJECT_SUMMARY normal lipid storage in cells. The gene encoding FIT2 is essential for the PR:PROJECT_SUMMARY viability of mice and worms. Whether FIT2 acts as an acyl-CoA diphosphatase in PR:PROJECT_SUMMARY vivo and how this activity affects liver, where the protein was discovered, are PR:PROJECT_SUMMARY unknown. Here, we report that hepatocyte-specific Fitm2 knockout (FIT2-LKO) mice PR:PROJECT_SUMMARY fed a chow diet exhibited elevated acyl-CoA levels, ER stress, and signs of PR:PROJECT_SUMMARY liver injury. These mice also had more triglycerides in their livers than PR:PROJECT_SUMMARY control littermates due, in part, to impaired secretion of triglyceride-rich PR:PROJECT_SUMMARY lipoproteins and reduced capacity for fatty acid oxidation. Challenging FIT2-LKO PR:PROJECT_SUMMARY mice with a high-fat diet worsened hepatic ER stress and liver injury, but PR:PROJECT_SUMMARY unexpectedly reversed the steatosis phenotype, similar to what is observed in PR:PROJECT_SUMMARY FIT2-deficient cells loaded with fatty acids. Our findings support the model PR:PROJECT_SUMMARY that FIT2 acts as an acyl-CoA diphosphatase in vivo and is crucial for normal PR:PROJECT_SUMMARY hepatocyte function and ER homeostasis in murine liver. PR:INSTITUTE Harvard University PR:DEPARTMENT Molecular Metabolism PR:LABORATORY FareseWalther's lab PR:LAST_NAME Bond PR:FIRST_NAME Laura PR:ADDRESS 655 Huntington Avenue, Building 1, Room 203, Boston, MA, 02115, USA PR:EMAIL laurabond44@gmail.com PR:PHONE +18573087183 #STUDY ST:STUDY_TITLE Fitm2 is required for ER homeostasis and normal function of murine liver ST:STUDY_SUMMARY The ER-resident protein fat-inducing transcript 2 (FIT2) catalyzes acyl-CoA ST:STUDY_SUMMARY cleavage in vitro and is required for endoplasmic reticulum (ER) homeostasis and ST:STUDY_SUMMARY normal lipid storage in cells. The gene encoding FIT2 is essential for the ST:STUDY_SUMMARY viability of mice and worms. Whether FIT2 acts as an acyl-CoA diphosphatase in ST:STUDY_SUMMARY vivo and how this activity affects liver, where the protein was discovered, are ST:STUDY_SUMMARY unknown. Here, we report that hepatocyte-specific Fitm2 knockout (FIT2-LKO) mice ST:STUDY_SUMMARY fed a chow diet exhibited elevated acyl-CoA levels, ER stress, and signs of ST:STUDY_SUMMARY liver injury. These mice also had more triglycerides in their livers than ST:STUDY_SUMMARY control littermates due, in part, to impaired secretion of triglyceride-rich ST:STUDY_SUMMARY lipoproteins and reduced capacity for fatty acid oxidation. Challenging FIT2-LKO ST:STUDY_SUMMARY mice with a high-fat diet worsened hepatic ER stress and liver injury, but ST:STUDY_SUMMARY unexpectedly reversed the steatosis phenotype, similar to what is observed in ST:STUDY_SUMMARY FIT2-deficient cells loaded with fatty acids. Our findings support the model ST:STUDY_SUMMARY that FIT2 acts as an acyl-CoA diphosphatase in vivo and is crucial for normal ST:STUDY_SUMMARY hepatocyte function and ER homeostasis in murine liver. ST:INSTITUTE Harvard School of Public Health ST:LAST_NAME Bond ST:FIRST_NAME Laura ST:ADDRESS 665 Huntington Ave., Building 2, 3rd Floor, Room 311 | Boston, MA 02115 ST:EMAIL laurabond44@gmail.com ST:PHONE 8573087183 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus #FACTORS #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 - Blank Genotype:Blank RAW_FILE_NAME=blank SUBJECT_SAMPLE_FACTORS - FIT2Flox_01 Genotype:flox RAW_FILE_NAME=FIT2Flox_01 SUBJECT_SAMPLE_FACTORS - FIT2Flox_02 Genotype:flox RAW_FILE_NAME=FIT2Flox_02 SUBJECT_SAMPLE_FACTORS - FIT2Flox_03 Genotype:flox RAW_FILE_NAME=FIT2Flox_03 SUBJECT_SAMPLE_FACTORS - FIT2Flox_04 Genotype:flox RAW_FILE_NAME=FIT2Flox_04 SUBJECT_SAMPLE_FACTORS - FIT2Flox_05 Genotype:flox RAW_FILE_NAME=FIT2Flox_05 SUBJECT_SAMPLE_FACTORS - FIT2-LKO_01 Genotype:Liver Knockout RAW_FILE_NAME=FIT2-LKO_01 SUBJECT_SAMPLE_FACTORS - FIT2-LKO_02 Genotype:Liver Knockout RAW_FILE_NAME=FIT2-LKO_02 SUBJECT_SAMPLE_FACTORS - FIT2-LKO_03 Genotype:Liver Knockout RAW_FILE_NAME=FIT2-LKO_03 SUBJECT_SAMPLE_FACTORS - FIT2-LKO_04 Genotype:Liver Knockout RAW_FILE_NAME=FIT2-LKO_04 SUBJECT_SAMPLE_FACTORS - FIT2-LKO_05 Genotype:Liver Knockout RAW_FILE_NAME=FIT2-LKO_05 SUBJECT_SAMPLE_FACTORS - FIT2-LKO_06 Genotype:Liver Knockout RAW_FILE_NAME=FIT2-LKO_06 #COLLECTION CO:COLLECTION_SUMMARY All mice fasted 2 hours prior to sacrifice. Mice were euthanized with isoflurane CO:COLLECTION_SUMMARY and tissues were collected. CO:SAMPLE_TYPE Liver #TREATMENT TR:TREATMENT_SUMMARY No treatment #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Liver (~100mg) was homogenized in 1 mL of PBS using a Bead Mill Homogenizer SP:SAMPLEPREP_SUMMARY (VWR). Lipids were extracted, according to the Folch method. SP:PROCESSING_STORAGE_CONDITIONS On ice SP:EXTRACT_STORAGE -80℃ #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Thermo Dionex Ultimate 3000 CH:COLUMN_NAME Thermo Accucore C30, 2.1 x 250 mm, 3 μm #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS Mass spectrometry data analysis was performed using LipidSearch version 4.1 SP MS:MS_COMMENTS (Thermo Fisher Scientific). The results were exported to R-Studio where quality MS:MS_COMMENTS control was performed using pairwise correlations between replicates, a MS:MS_COMMENTS principal component analysis comparing sample groups, as well as retention time MS:MS_COMMENTS plot analysis to verify elution clustering within lipid classes. All identified MS:MS_COMMENTS lipids were included for subsequent analyses if they fulfilled the following MS:MS_COMMENTS LipidSearch-based criteria: 1) reject equal to zero, 2) main grade A OR main MS:MS_COMMENTS grade B AND APvalue<0.01 for at least three replicates, and 3) no missing values MS:MS_COMMENTS across all samples. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Area MS_METABOLITE_DATA_START Samples Blank FIT2Flox_01 FIT2Flox_02 FIT2Flox_03 FIT2Flox_04 FIT2Flox_05 FIT2-LKO_01 FIT2-LKO_02 FIT2-LKO_03 FIT2-LKO_04 FIT2-LKO_05 FIT2-LKO_06 Factors Genotype:Blank Genotype:flox Genotype:flox Genotype:flox Genotype:flox Genotype:flox Genotype:Liver Knockout Genotype:Liver Knockout Genotype:Liver Knockout Genotype:Liver Knockout Genotype:Liver Knockout Genotype:Liver Knockout Cer(d17:0+pO/22:0) 0 5210000 4870000 4030000 4690000 4340000 5250000 5370000 4620000 5020000 5110000 4820000 Cer(d17:0+pO/23:0) 0 6190000 4270000 4090000 4970000 5970000 5260000 4550000 6500000 4630000 5510000 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4920000000 4930000000 5410000000 8120000000 4450000000 TG(19:1/18:1/18:1) 62500 66800000 345000000 732000000 302000000 325000000 1040000000 1830000000 2010000000 1910000000 2880000000 1710000000 TG(19:1/18:1/18:2) 56200 416000000 428000000 846000000 547000000 429000000 1170000000 1820000000 1810000000 2340000000 2630000000 2270000000 TG(19:1/18:1/20:3) 1060000 48500000 55600000 62600000 67700000 54000000 149000000 155000000 169000000 174000000 198000000 181000000 TG(19:1/18:1/22:5) 0 29300000 16600000 24800000 18200000 51000000 85000000 71600000 48800000 86100000 98400000 69400000 TG(19:1/18:2/18:2) 76300 283000000 211000000 322000000 312000000 261000000 626000000 577000000 570000000 607000000 610000000 585000000 TG(19:1/18:2/18:3) 0 177000000 162000000 127000000 199000000 183000000 250000000 195000000 239000000 271000000 220000000 86300000 TG(19:1/18:2/22:5) 47200 24600000 42500000 50900000 58700000 69900000 116000000 26300000 84300000 97700000 98600000 74900000 TG(19:1/18:2/22:6) 0 39000000 29300000 29600000 6990000 37600 29800000 29900000 6890000 49600000 73600000 50800000 TG(20:0/16:0/18:1) 601000 631000000 1610000000 2160000000 1140000000 1160000000 3290000000 7870000000 4300000000 6030000000 7230000000 7720000000 TG(20:0/16:0/18:2) 1060000 993000000 2230000000 2250000000 1730000000 1530000000 3580000000 6830000000 4520000000 5740000000 7000000000 7090000000 TG(20:0/18:1/18:1) 70900 720000000 1630000000 2690000000 1300000000 969000000 5450000000 14200000000 6860000000 6750000000 11400000000 8500000000 TG(20:0/18:1/18:2) 186000 1400000000 3770000000 4220000000 2130000000 2220000000 9360000000 16300000000 11300000000 16000000000 16300000000 16000000000 TG(20:0/18:1/22:5) 0 77700000 116000000 131000000 102000000 93100000 315000000 434000000 352000000 506000000 427000000 425000000 TG(20:0/18:1/22:6) 0 150000000 255000000 235000000 222000000 170000000 546000000 686000000 418000000 722000000 649000000 656000000 TG(20:0/18:2/18:3) 51200 1290000000 1360000000 1650000000 1730000000 1440000000 2220000000 2510000000 2100000000 1950000000 1970000000 1930000000 TG(20:0/18:2/22:5) 0 206000000 106000000 93500000 164000000 97200000 238000000 209000000 167000000 216000000 229000000 226000000 TG(20:0e/16:0/16:0) 0 27700000 20800000 17800000 17200000 18600000 23800000 37500000 33100000 24400000 32500000 28400000 TG(20:0e/16:0/18:1) 53100 50000000 38800000 32800000 31100000 31200000 49600000 85300000 73500000 57000000 78400000 63900000 TG(20:0e/16:0/18:2) 0 55600000 30100000 27300000 29300000 28700000 48200000 69800000 66900000 58400000 69600000 62300000 TG(20:0e/18:1/18:1) 0 21800000 18600000 15800000 15200000 15000000 38400000 64100000 51500000 48300000 71900000 40200000 TG(20:0p/16:0/16:0) 58900 119000000 79200000 91700000 75000000 82200000 85500000 143000000 129000000 102000000 140000000 103000000 TG(20:1/18:1/18:1) 88600 1850000000 2320000000 5120000000 2050000000 1730000000 11500000000 15500000000 16700000000 17300000000 20100000000 12900000000 TG(20:1/18:1/18:2) 50400 3860000000 3990000000 6300000000 4390000000 3150000000 13500000000 12700000000 12700000000 14800000000 14400000000 11100000000 TG(20:1/18:1/20:1) 0 220000000 394000000 684000000 282000000 254000000 1850000000 3530000000 2520000000 3960000000 4370000000 3030000000 TG(20:1/18:1/20:3) 27900 482000000 637000000 830000000 674000000 467000000 2480000000 2710000000 2300000000 3220000000 3150000000 2710000000 TG(20:1/18:1/22:4) 0 83900000 114000000 160000000 103000000 91300000 462000000 554000000 416000000 656000000 633000000 684000000 TG(20:1/18:1/22:5) 0 581000000 382000000 540000000 771000000 161000000 1150000000 1460000000 1420000000 1760000000 1660000000 1420000000 TG(20:1/18:1/22:6) 0 423000000 483000000 694000000 458000000 332000000 2560000000 1610000000 2070000000 2610000000 2030000000 1640000000 TG(20:1/18:2/22:5) 0 554000000 137000000 194000000 202000000 404000000 442000000 1250000000 304000000 408000000 352000000 1310000000 TG(20:1/18:2/22:6) 0 672000000 806000000 1070000000 1100000000 828000000 2200000000 1390000000 1710000000 2090000000 1550000000 1360000000 TG(20:1/20:3/22:5) 0 42600000 28200000 31400000 29500000 29100000 98000000 56100000 56700000 70100000 53700000 48900000 TG(20:1/22:5/22:5) 0 13600000 3610000 12900000 7540000 9230000 45400000 22100000 41600000 47900000 34700000 33000000 TG(20:1/22:6/22:6) 60200 23400000 2070000 17300000 15200000 15700000 48700000 26800000 45100000 38800000 54400000 43300000 TG(20:3/22:5/22:6) 0 50600000 43200000 57300000 49900000 38800000 117000000 70900000 91600000 106000000 85100000 94100000 TG(20:3/22:6/22:6) 0 35400000 9520000 14500000 26400000 21600000 26400000 65300000 61400000 81300000 19300000 71700000 TG(20:4/22:6/22:6) 0 16200000 14900000 22700000 21900000 18900000 20800000 22100000 22000000 24800000 27400000 15400000 TG(20:5/14:2/18:2) 0 11500000 13300000 19900000 37200000 28500000 14500000 25400000 8990000 17500000 17700000 17900000 TG(20:5/17:1/22:6) 0 16100000 12800000 20300000 21100000 15100000 19200000 22400000 19500000 85500000 23700000 76400000 TG(20:5/18:2/22:6) 0 1000000000 750000000 1110000000 1290000000 1040000000 1100000000 883000000 757000000 961000000 995000000 841000000 TG(20:5/22:6/22:6) 0 12200000 12300000 17600000 18100000 15900000 17900000 18000000 14600000 19200000 22800000 16100000 TG(22:0/18:2/22:5) 10600000 13300000 24500000 22700000 16800000 15600000 51200000 34600000 35100000 42800000 44600000 51300000 TG(22:0/18:2/22:6) 0 16100000 21200000 16400000 24500000 13800000 23900000 25100000 14400000 24700000 20100000 30000000 TG(22:1/18:2/22:5) 0 35200000 32000000 35800000 29000000 35100000 84800000 46300000 58900000 65700000 47300000 66000000 TG(22:1/18:2/22:6) 0 89800000 93700000 105000000 104000000 77300000 261000000 248000000 157000000 265000000 222000000 275000000 TG(22:4/18:2/22:6) 0 175000000 178000000 236000000 239000000 167000000 546000000 321000000 353000000 408000000 389000000 263000000 TG(22:5/17:1/20:4) 0 31700000 24300000 31700000 27000000 30200000 58700000 17700000 64100000 51000000 65900000 34300000 TG(22:5/18:2/18:2) 0 1430000000 1350000000 1670000000 1880000000 1170000000 3720000000 2420000000 2990000000 3230000000 3380000000 2610000000 TG(22:5/18:2/22:6) 0 413000000 305000000 404000000 440000000 308000000 988000000 684000000 729000000 882000000 838000000 671000000 TG(22:5/22:5/22:6) 0 2490000 3560000 4440000 2330000 1700000 20200000 16300000 20600000 19000000 21000000 13000000 TG(22:5/22:6/22:6) 0 13800000 12000000 20700000 15800000 9620000 66700000 37600000 52600000 59200000 59000000 39000000 TG(22:6/17:1/22:6) 0 34500000 23900000 35000000 41400000 27100000 66800000 50900000 51500000 57500000 65300000 48600000 TG(22:6/22:6/22:6) 0 17800000 14200000 22300000 16900000 15100000 41900000 29400000 30800000 36000000 37000000 24700000 TG(24:1/18:2/18:2) 369000 92300000 117000000 103000000 123000000 91000000 278000000 401000000 242000000 384000000 458000000 359000000 TG(24:1/18:2/22:6) 0 11800000 10700000 11300000 15000000 12000000 25000000 21200000 15300000 31000000 19100000 30700000 TG(26:0/16:0/18:1) 0 7700000 12800000 15000000 11900000 14100000 22900000 44300000 35600000 43300000 49900000 57600000 TG(26:0/16:0/18:2) 55100 7500000 11500000 12100000 10800000 14500000 14600000 27400000 15400000 17400000 26000000 35800000 TG(26:0/18:1/18:1) 0 7950000 11400000 10900000 9250000 9780000 21000000 41700000 22400000 48800000 54300000 54100000 TG(26:0/18:1/18:2) 56600 8270000 12500000 9070000 12100000 10700000 19000000 30900000 21200000 31000000 28900000 40000000 TG(26:1/18:1/18:1) 44600 6010000 8760000 9910000 6060000 8490000 22400000 33800000 28900000 41300000 44900000 37200000 TG(26:1/18:1/18:2) 1840000 11200000 15100000 19100000 14100000 16100000 28200000 55700000 32700000 50700000 51300000 47200000 TG(26:1/18:1/24:2) 0 3660000 6070000 11400000 5960000 6860000 9580000 8710000 29500000 26400000 35000000 23500000 TG(26:1/18:2/18:2) 33900 4600000 7890000 15100000 5900000 7330000 7990000 17200000 9990000 13700000 14400000 20200000 TG(28:0/18:1/24:2) 0 5530000 6270000 16000000 4420000 10000000 12300000 7430000 47500000 24600000 41600000 35300000 TG(28:1/18:1/24:2) 0 8200000 9150000 17400000 7870000 12800000 17800000 8280000 40300000 30100000 41400000 37800000 TG(30:0/18:1/18:1) 0 2790000 7780000 16400000 3890000 5740000 4010000 6550000 20400000 17700000 26500000 18200000 TG(30:0/18:1/20:1) 0 4380000 7640000 25000000 4540000 10200000 8380000 6550000 34300000 19800000 33100000 31500000 TG(30:1/18:1/20:1) 0 5300000 8170000 17700000 6020000 5200000 6900000 6810000 31000000 30000000 38600000 31300000 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Cer(d17:0+pO/22:0) Cer(d17:0+pO/23:0) Cer(d17:0+pO/24:0) Cer(d17:0+pO/24:1) Cer(d17:1/22:0) Cer(d17:1/24:1) Cer(d18:0/22:0) Cer(d18:0/23:0) Cer(d18:0/24:0) Cer(d18:0+pO/22:0) Cer(d18:0+pO/23:0) Cer(d18:0+pO/38:3) Cer(d18:0+pO/40:4) Cer(d18:1/16:0) Cer(d18:1/17:0) Cer(d18:1/18:0) Cer(d18:1/20:0) Cer(d18:1/22:0) Cer(d18:1/22:1) Cer(d18:1/23:0) Cer(d18:1/24:0) Cer(d18:1/24:1) Cer(d18:1/24:2) Cer(d18:1/25:0) Cer(d18:1/25:1) Cer(d18:1/26:0) Cer(d18:2/20:0) Cer(d18:2/22:0) Cer(d18:2/23:0) Cer(d18:2/24:0) Cer(d18:2/24:1) Cer(d20:2/18:0+O) CerG1(d18:1/12:0) CerG1(d18:1/16:0) CerG1(d18:1/20:0) CerG1(d18:1/22:0) CerG1(d18:1/23:0) CerG1(d18:1/24:0) CerG1(d18:1/24:1) ChE(16:0) ChE(16:1) ChE(17:1) ChE(18:0) ChE(18:1) ChE(18:2) ChE(18:3) ChE(20:2) ChE(20:3) ChE(20:4) ChE(20:5) ChE(22:4) ChE(22:5) ChE(22:6) CL(17:1/18:1/18:2/18:2) CL(17:1/18:2/18:2/18:2) CL(18:1/16:0/16:0/18:1) CL(18:1/16:0/16:1/18:2) CL(18:1/18:1/18:1/16:0) CL(18:2/16:0/16:0/18:1) CL(18:2/16:0/18:2/18:1) CL(18:2/16:0/18:2/18:2) CL(18:2/16:1/16:1/18:1) CL(18:2/16:1/16:1/18:2) CL(18:2/16:1/18:2/18:1) CL(18:2/16:1/18:2/18:2) CL(18:2/16:1/22:6/16:1) CL(18:2/18:1/16:0/18:1) CL(18:2/18:1/18:1/18:2) CL(18:2/18:1/22:5/18:2) CL(18:2/18:2/18:1/16:0) CL(18:2/18:2/18:2/18:1) CL(18:2/18:2/18:2/18:2) CL(18:2/18:2/18:2/20:3) CL(18:2/18:2/18:2/22:2) CL(18:2/18:2/18:2/22:5) CL(18:2/18:2/18:2/22:6) CL(18:2/20:3/18:2/20:3) CL(18:2/20:3/22:5/18:2) CL(18:2/20:3/22:6/18:2) CL(18:2/20:4/18:1/18:2) CL(18:2/22:6/18:2/20:3) CL(18:3/16:1/18:2/18:2) CL(18:3/18:2/22:6/16:1) CL(18:4/18:2/16:1/18:2) CL(18:4/18:2/18:2/18:2) CL(19:0/18:0/18:1/22:6) CL(20:2/16:0/22:6/22:6) CL(20:2/18:2/18:2/18:1) CL(20:2/18:2/18:2/18:2) CL(20:5/18:2/18:1/18:2) CL(20:5/18:2/18:2/18:2) CL(21:0/16:0/20:3/22:6) CL(21:0/16:0/22:5/22:6) CL(22:2/18:0/20:4/20:4) CL(22:4/20:1/18:0/20:4) CL(22:6/16:0/20:0/22:6) CL(22:6/16:1/22:6/18:2) CL(22:6/18:1/20:0/22:6) CL(23:0/18:0/18:0/20:4) DG(12:0/18:2) DG(14:0/20:4) DG(14:0/22:6) DG(15:0/18:1) DG(15:0/18:2) DG(16:0/14:0) DG(16:0/16:0) DG(16:0/16:1) DG(16:0/18:1) DG(16:0/18:2) DG(16:0/18:3) DG(16:0/20:4) DG(16:0/20:5) DG(16:0/22:6) DG(16:1/14:0) DG(16:1/16:1) DG(16:1/18:1) DG(16:1/18:2) DG(16:1/18:3) DG(16:1/20:4) DG(16:1/20:5) DG(16:1/22:6) DG(17:0/20:4) DG(17:1/18:1) DG(17:1/18:2) DG(18:0/16:0) DG(18:0/18:0) DG(18:0/18:1) DG(18:0/18:2) DG(18:0/20:3) DG(18:0/20:4) DG(18:0/22:4) DG(18:0/22:6) DG(18:1/18:1) DG(18:1/18:2) DG(18:1/18:3) DG(18:1/20:3) DG(18:1/20:4) DG(18:1/20:5) DG(18:1/22:0) DG(18:1/22:1) DG(18:1/22:5) DG(18:1/22:6) DG(18:2/18:2) DG(18:2/20:4) DG(18:2/22:6) DG(18:3/18:2) DG(18:3/18:3) DG(18:3/20:5) DG(18:3/22:6) DG(19:1/18:1) DG(19:1/18:2) DG(20:0/18:1) DG(20:0/18:2) DG(20:0/22:6) DG(20:1/18:1) DG(20:1/18:2) DG(20:1/22:6) DG(20:3/22:5) DG(20:4/22:6) DG(20:5/18:2) DG(22:1/18:2) DG(22:5/18:2) DG(22:5/22:6) DG(22:6/22:6) So(d14:0+pO) So(d16:0+pO) So(d18:0) So(d18:0+pO) So(d18:1) So(d20:0+pO) TG(10:0/18:1/22:6) TG(10:0/18:2/18:2) TG(11:0/16:0/18:2) TG(11:0/18:2/18:2) TG(12:0/18:2/20:5) TG(14:0/18:2/18:3) TG(14:0/20:5/22:6) TG(15:0/16:0/16:0) TG(15:0/16:0/16:1) TG(15:0/16:0/18:1) TG(15:0/16:0/18:2) TG(15:0/16:1/16:1) TG(15:0/16:1/18:3) TG(15:0/18:1/20:5) TG(15:0/18:1/22:6) TG(15:0/18:2/18:3) TG(15:0/18:2/20:5) TG(15:0/18:2/22:6) TG(15:0/18:3/18:3) TG(15:0/18:3/22:6) TG(15:0/20:5/22:6) TG(15:0/22:5/22:6) TG(15:0/22:6/22:6) TG(16:0/11:1/18:2) TG(16:0/12:0/14:0) TG(16:0/14:0/14:0) TG(16:0/14:0/16:0) TG(16:0/14:0/16:1) TG(16:0/14:0/18:1) TG(16:0/14:1/20:5) TG(16:0/16:0/16:0) TG(16:0/16:0/17:0) TG(16:0/16:0/18:1) TG(16:0/16:0/20:5) TG(16:0/16:0/22:6) TG(16:0/16:0/24:0) TG(16:0/16:1/18:1) TG(16:0/16:1/18:3) TG(16:0/17:0/18:1) TG(16:0/17:0/18:2) TG(16:0/17:1/20:4) TG(16:0/18:1/18:1) TG(16:0/18:1/18:2) TG(16:0/18:1/18:3) TG(16:0/18:1/19:0) TG(16:0/18:1/20:1) TG(16:0/18:1/21:0) TG(16:0/18:1/22:0) TG(16:0/18:1/22:5) TG(16:0/18:1/22:6) TG(16:0/18:1/23:0) TG(16:0/18:1/24:0) TG(16:0/18:2/18:2) TG(16:0/18:2/18:3) TG(16:0/18:2/19:0) TG(16:0/18:2/20:3) TG(16:0/18:2/20:5) TG(16:0/18:2/22:0) TG(16:0/18:2/22:6) TG(16:0/18:2/24:0) TG(16:0/18:3/22:6) TG(16:0/20:5/22:6) TG(16:0/22:6/22:6) TG(16:0/8:0/18:1) TG(16:0/8:0/22:6) TG(16:0/9:0/18:2) TG(16:0e/16:0/16:0) TG(16:0e/16:0/20:4) TG(16:0e/16:0/22:4) TG(16:0e/18:1/18:2) TG(16:0e/18:1/22:5) TG(16:1/10:0/18:2) TG(16:1/12:0/16:1) TG(16:1/13:0/18:2) TG(16:1/14:0/14:0) TG(16:1/14:0/16:1) TG(16:1/14:0/18:1) TG(16:1/14:0/22:6) TG(16:1/14:1/16:1) TG(16:1/14:1/18:2) TG(16:1/16:1/16:1) TG(16:1/16:1/17:1) TG(16:1/16:1/18:1) TG(16:1/16:1/18:2) TG(16:1/16:1/22:6) TG(16:1/17:1/18:2) TG(16:1/18:2/18:3) TG(16:1/18:2/22:5) TG(17:0/18:1/18:1) TG(17:0/18:1/20:4) TG(17:0/18:1/20:5) TG(17:0/18:1/22:5) TG(17:0/18:1/22:6) TG(17:0/18:2/22:6) TG(18:0/16:0/16:0) TG(18:0/16:0/17:0) TG(18:0/16:0/18:0) TG(18:0/16:0/18:1) TG(18:0/18:0/18:0) TG(18:0/18:1/22:4) TG(18:0/18:1/22:5) TG(18:0/20:1/20:4) TG(18:0/20:3/20:4) TG(18:0/20:4/22:6) TG(18:0/22:5/22:6) TG(18:0e/16:0/16:0) TG(18:0e/16:0/18:2) TG(18:0e/16:0/18:3) TG(18:0e/16:0/22:4) TG(18:0e/18:0/22:5) TG(18:0p/16:0/16:0) TG(18:0p/16:0/16:1) TG(18:0p/16:0/18:1) TG(18:0p/16:0/20:4) TG(18:0p/16:0/22:5) TG(18:0p/18:0/20:4) TG(18:0p/18:0/22:5) TG(18:0p/18:1/18:1) TG(18:0p/18:1/18:2) TG(18:1/17:1/18:2) TG(18:1/17:1/22:5) TG(18:1/18:1/18:1) TG(18:1/18:1/18:2) TG(18:1/18:1/20:4) TG(18:1/18:1/21:0) TG(18:1/18:1/22:0) TG(18:1/18:1/22:5) TG(18:1/18:1/22:6) TG(18:1/18:1/23:0) TG(18:1/18:1/23:1) TG(18:1/18:1/24:0) TG(18:1/18:1/24:1) TG(18:1/18:2/18:2) TG(18:1/18:2/18:3) TG(18:1/18:2/20:2) TG(18:1/18:2/20:4) TG(18:1/18:2/21:0) TG(18:1/18:2/21:1) TG(18:1/18:2/22:0) TG(18:1/18:2/22:1) TG(18:1/18:2/22:3) TG(18:1/18:2/22:5) TG(18:1/18:2/22:6) TG(18:1/18:2/23:0) TG(18:1/18:2/23:1) TG(18:1/18:2/24:1) TG(18:1/20:3/22:5) TG(18:1/20:3/22:6) TG(18:1/20:3/24:0) TG(18:1/20:4/22:5) TG(18:1/20:4/22:6) TG(18:1/20:5/22:6) TG(18:1/22:0/22:5) TG(18:1/22:0/22:6) TG(18:1/22:1/22:4) TG(18:1/22:1/22:5) TG(18:1/22:1/22:6) TG(18:1/22:4/22:5) TG(18:1/22:5/22:5) TG(18:1/22:5/22:6) TG(18:1/22:6/22:6) TG(18:1/22:6/24:0) TG(18:1/22:6/24:1) TG(18:2/13:0/22:6) TG(18:2/17:1/18:2) TG(18:2/17:1/22:6) TG(18:2/18:2/23:1) TG(18:2/22:6/22:6) TG(18:3/12:1/18:2) TG(18:3/14:1/18:2) TG(18:3/17:1/18:2) TG(18:3/17:1/22:6) TG(18:3/18:2/18:2) TG(18:3/18:2/18:3) TG(18:3/18:2/20:4) TG(18:3/18:2/20:5) TG(18:3/18:2/22:5) TG(18:3/18:2/22:6) TG(18:3/18:3/20:5) TG(18:3/18:3/22:6) TG(18:3/20:5/20:5) TG(18:3/20:5/22:6) TG(18:3/22:6/22:6) TG(18:4/16:0/16:1) TG(18:4/16:1/18:2) TG(18:4/16:1/18:3) TG(18:4/18:2/18:3) TG(18:4/18:3/18:3) TG(19:0/18:1/18:1) TG(19:0/18:1/20:3) TG(19:0/18:1/20:4) TG(19:0/18:1/22:5) TG(19:0/18:2/20:3) TG(19:0/18:2/22:6) TG(19:1/17:1/17:1) TG(19:1/18:1/18:1) TG(19:1/18:1/18:2) TG(19:1/18:1/20:3) TG(19:1/18:1/22:5) TG(19:1/18:2/18:2) TG(19:1/18:2/18:3) TG(19:1/18:2/22:5) TG(19:1/18:2/22:6) TG(20:0/16:0/18:1) TG(20:0/16:0/18:2) TG(20:0/18:1/18:1) TG(20:0/18:1/18:2) TG(20:0/18:1/22:5) TG(20:0/18:1/22:6) TG(20:0/18:2/18:3) TG(20:0/18:2/22:5) TG(20:0e/16:0/16:0) TG(20:0e/16:0/18:1) TG(20:0e/16:0/18:2) TG(20:0e/18:1/18:1) TG(20:0p/16:0/16:0) TG(20:1/18:1/18:1) TG(20:1/18:1/18:2) TG(20:1/18:1/20:1) TG(20:1/18:1/20:3) TG(20:1/18:1/22:4) TG(20:1/18:1/22:5) TG(20:1/18:1/22:6) TG(20:1/18:2/22:5) TG(20:1/18:2/22:6) TG(20:1/20:3/22:5) TG(20:1/22:5/22:5) TG(20:1/22:6/22:6) TG(20:3/22:5/22:6) TG(20:3/22:6/22:6) TG(20:4/22:6/22:6) TG(20:5/14:2/18:2) TG(20:5/17:1/22:6) TG(20:5/18:2/22:6) TG(20:5/22:6/22:6) TG(22:0/18:2/22:5) TG(22:0/18:2/22:6) TG(22:1/18:2/22:5) TG(22:1/18:2/22:6) TG(22:4/18:2/22:6) TG(22:5/17:1/20:4) TG(22:5/18:2/18:2) TG(22:5/18:2/22:6) TG(22:5/22:5/22:6) TG(22:5/22:6/22:6) TG(22:6/17:1/22:6) TG(22:6/22:6/22:6) TG(24:1/18:2/18:2) TG(24:1/18:2/22:6) TG(26:0/16:0/18:1) TG(26:0/16:0/18:2) TG(26:0/18:1/18:1) TG(26:0/18:1/18:2) TG(26:1/18:1/18:1) TG(26:1/18:1/18:2) TG(26:1/18:1/24:2) TG(26:1/18:2/18:2) TG(28:0/18:1/24:2) TG(28:1/18:1/24:2) TG(30:0/18:1/18:1) TG(30:0/18:1/20:1) TG(30:1/18:1/20:1) METABOLITES_END #END