#METABOLOMICS WORKBENCH Jingbo_20250928_230926 DATATRACK_ID:6499 STUDY_ID:ST004258 ANALYSIS_ID:AN007086 PROJECT_ID:PR002686 VERSION 1 CREATED_ON October 6, 2025, 10:21 am #PROJECT PR:PROJECT_TITLE Mitochondria-localized MLKL channel function drives proinflammatory response PR:PROJECT_TITLE independent of necroptosis PR:PROJECT_SUMMARY Mixed lineage kinase domain-like protein (MLKL) has been well-established as an PR:PROJECT_SUMMARY executor of necroptosis. However, its necroptosis independent functions have PR:PROJECT_SUMMARY recently garnered attention, particularly its involvement in non-necroptotic PR:PROJECT_SUMMARY pathological processes. Here, we found that MLKL expression was upregulated in PR:PROJECT_SUMMARY macrophages during inflammation. Our proteomic and functional assays revealed PR:PROJECT_SUMMARY that, despite the absence of necroptosis, metabolic dysregulation and PR:PROJECT_SUMMARY inflammatory responses were still MLKL-dependent. These phenomena were notably PR:PROJECT_SUMMARY suppressed in MLKL-deficient macrophages. Further investigation showed that MLKL PR:PROJECT_SUMMARY translocates to the mitochondrial membrane during inflammation, where its PR:PROJECT_SUMMARY interaction with cardiolipin enhances channel activity, resulting in PR:PROJECT_SUMMARY mitochondrial dysfunction and an amplified inflammatory response. In both in PR:PROJECT_SUMMARY vitro and in vivo models, MLKL deficiency restored mitochondrial function, PR:PROJECT_SUMMARY relieved inflammation, reduced tissue damage. These findings establish MLKL PR:PROJECT_SUMMARY channel as a crucial regulator of macrophage inflammation, independent of its PR:PROJECT_SUMMARY role in necroptosis, and highlight its potential as a therapeutic target in PR:PROJECT_SUMMARY inflammation-associated diseases such as sepsis. PR:INSTITUTE Fudan University PR:DEPARTMENT Institutes of Biomedical Sciences PR:LABORATORY Systems biology Laboratory PR:LAST_NAME Qie PR:FIRST_NAME Jingbo PR:ADDRESS Institutes of Biomedical Sciences, Fudan University, No. 131, Dong 'an Road, PR:ADDRESS Shanghai 200032, China PR:EMAIL jingboqie@fudan.edu.cn PR:PHONE 17717890408 #STUDY ST:STUDY_TITLE Mitochondria-localized MLKL channel function drives proinflammatory response ST:STUDY_TITLE independent of necroptosis ST:STUDY_SUMMARY Experiment design: BMDMs were isolated from the bone marrow of Mlkl-/-/Mlkl+/+ ST:STUDY_SUMMARY and Ripk3-/-/Ripk3+/+ mouse femurs and tibias and cultured in DMEM medium ST:STUDY_SUMMARY (Gibco, C11995500BT, USA) containing 25 ng/mL M-CSF (meilunbio, MA0608, China) ST:STUDY_SUMMARY and 10% FBS (Gibco, A5669701, USA) for 7 days, with a medium change at 3.5 days. ST:STUDY_SUMMARY After successful differentiation, cells were incubated with PBS (meilunbio, ST:STUDY_SUMMARY MA0015, China) or 100 ng/mL LPS (sigma, L2630, USA) for 24 hours. Each ST:STUDY_SUMMARY experimental group of three biological replicates was performed, with each ST:STUDY_SUMMARY biological replicate undergoing two technical replicates. As a result, we ST:STUDY_SUMMARY discovered that deletion of either MLKL or RIPK3 resulted in a comparable ST:STUDY_SUMMARY reduction of key inflammatory metabolites, including succinate, fumarate, ST:STUDY_SUMMARY lactate, and citrate. Considering the effects of MLKL and RIPK3 on ROS, MMP, the ST:STUDY_SUMMARY NF-κB signaling pathway, and metabolomic profiles, we speculate that although ST:STUDY_SUMMARY MLKL functions in a RIPK3-independent manner, deficiencies in either MLKL or ST:STUDY_SUMMARY RIPK3 can influence proinflammatory responses and metabolic reprogramming. These ST:STUDY_SUMMARY processes may be interconnected through multiple feedback regulatory mechanisms. ST:INSTITUTE Fudan University ST:DEPARTMENT Institutes of Biomedical Sciences ST:LABORATORY Systems biology Laboratory ST:LAST_NAME Qie ST:FIRST_NAME Jingbo ST:ADDRESS Institutes of Biomedical Sciences, Fudan University, No. 131, Dong 'an Road, ST:ADDRESS Shanghai 200032, China ST:EMAIL jingboqie@fudan.edu.cn ST:PHONE 17717890408 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:GENOTYPE_STRAIN C57BL/6J SU:AGE_OR_AGE_RANGE 8-10 weeks SU:GENDER Male SU:CELL_STRAIN_DETAILS Bone-marrow derived macrophages (BMDM) SU:CELL_PRIMARY_IMMORTALIZED Primary Cells SU:CELL_PASSAGE_NUMBER 1 SU:CELL_COUNTS 1e6 #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 - wt1 pbs1-1 experimental factors:ctl1 | Sample source:wt1-mlkl-ko-littermate RAW_FILE_NAME(raw data file names)=wt1 pbs1-1.D SUBJECT_SAMPLE_FACTORS - wt1 pbs1-2 experimental factors:ctl1 | Sample source:wt1-mlkl-ko-littermate RAW_FILE_NAME(raw data file names)=wt1 pbs1-2.D SUBJECT_SAMPLE_FACTORS - wt1 pbs2-1 experimental factors:ctl1 | Sample source:wt1-mlkl-ko-littermate RAW_FILE_NAME(raw data file names)=wt1 pbs2-1.D SUBJECT_SAMPLE_FACTORS - wt1 pbs2-2 experimental factors:ctl1 | Sample source:wt1-mlkl-ko-littermate RAW_FILE_NAME(raw data file names)=wt1 pbs2-2.D SUBJECT_SAMPLE_FACTORS - wt1 pbs3-1 experimental factors:ctl1 | Sample source:wt1-mlkl-ko-littermate RAW_FILE_NAME(raw data file names)=wt1 pbs3-1.D SUBJECT_SAMPLE_FACTORS - wt1 pbs3-2 experimental factors:ctl1 | Sample source:wt1-mlkl-ko-littermate RAW_FILE_NAME(raw data file names)=wt1 pbs3-2.D SUBJECT_SAMPLE_FACTORS - wt1 lps1-1 experimental factors:lps | Sample source:wt1-mlkl-ko-littermate RAW_FILE_NAME(raw data file names)=wt1 lps1-1.D SUBJECT_SAMPLE_FACTORS - wt1 lps1-2 experimental factors:lps | Sample source:wt1-mlkl-ko-littermate RAW_FILE_NAME(raw data file names)=wt1 lps1-2.D SUBJECT_SAMPLE_FACTORS - wt1 lps2-1 experimental factors:lps | Sample source:wt1-mlkl-ko-littermate RAW_FILE_NAME(raw data file names)=wt1 lps2-1.D SUBJECT_SAMPLE_FACTORS - wt1 lps2-2 experimental factors:lps | Sample source:wt1-mlkl-ko-littermate RAW_FILE_NAME(raw data file names)=wt1 lps2-2.D SUBJECT_SAMPLE_FACTORS - wt1 lps3-1 experimental factors:lps | Sample source:wt1-mlkl-ko-littermate RAW_FILE_NAME(raw data file names)=wt1 lps3-1.D SUBJECT_SAMPLE_FACTORS - wt1 lps3-2 experimental factors:lps | Sample source:wt1-mlkl-ko-littermate RAW_FILE_NAME(raw data file names)=wt1 lps3-2.D SUBJECT_SAMPLE_FACTORS - mlkl-ko pbs1-1 experimental factors:ctl1 | Sample source:mlkl-ko RAW_FILE_NAME(raw data file names)=mlkl-ko pbs1-1.D SUBJECT_SAMPLE_FACTORS - mlkl-ko pbs1-2 experimental factors:ctl1 | Sample source:mlkl-ko RAW_FILE_NAME(raw data file names)=mlkl-ko pbs1-2.D SUBJECT_SAMPLE_FACTORS - mlkl-ko pbs2-1 experimental factors:ctl1 | Sample source:mlkl-ko RAW_FILE_NAME(raw data file names)=mlkl-ko pbs2-1.D SUBJECT_SAMPLE_FACTORS - mlkl-ko pbs2-2 experimental factors:ctl1 | Sample source:mlkl-ko RAW_FILE_NAME(raw data file names)=mlkl-ko pbs2-2.D SUBJECT_SAMPLE_FACTORS - mlkl-ko pbs3-1 experimental factors:ctl1 | Sample source:mlkl-ko RAW_FILE_NAME(raw data file names)=mlkl-ko pbs3-1.D SUBJECT_SAMPLE_FACTORS - mlkl-ko pbs3-2 experimental factors:ctl1 | Sample source:mlkl-ko RAW_FILE_NAME(raw data file names)=mlkl-ko pbs3-2.D SUBJECT_SAMPLE_FACTORS - mlkl-ko lps1-1 experimental factors:lps | Sample source:mlkl-ko RAW_FILE_NAME(raw data file names)=mlkl-ko lps1-1.D SUBJECT_SAMPLE_FACTORS - mlkl-ko lps1-2 experimental factors:lps | Sample source:mlkl-ko RAW_FILE_NAME(raw data file names)=mlkl-ko lps1-2.D SUBJECT_SAMPLE_FACTORS - mlkl-ko lps2-1 experimental factors:lps | Sample source:mlkl-ko RAW_FILE_NAME(raw data file names)=mlkl-ko lps2-1.D SUBJECT_SAMPLE_FACTORS - mlkl-ko lps2-2 experimental factors:lps | Sample source:mlkl-ko RAW_FILE_NAME(raw data file names)=mlkl-ko lps2-2.D SUBJECT_SAMPLE_FACTORS - mlkl-ko lps3-1 experimental factors:lps | Sample source:mlkl-ko RAW_FILE_NAME(raw data file names)=mlkl-ko lps3-1.D SUBJECT_SAMPLE_FACTORS - mlkl-ko lps3-2 experimental factors:lps | Sample source:mlkl-ko RAW_FILE_NAME(raw data file names)=mlkl-ko lps3-2.D SUBJECT_SAMPLE_FACTORS - wt2 pbs1-1 experimental factors:ctl1 | Sample source:wt2-ripk3-ko-littermate RAW_FILE_NAME(raw data file names)=wt2 pbs1-1.D SUBJECT_SAMPLE_FACTORS - wt2 pbs1-2 experimental factors:ctl1 | Sample source:wt2-ripk3-ko-littermate RAW_FILE_NAME(raw data file names)=wt2 pbs1-2.D SUBJECT_SAMPLE_FACTORS - wt2 pbs2-1 experimental factors:ctl1 | Sample source:wt2-ripk3-ko-littermate RAW_FILE_NAME(raw data file names)=wt2 pbs2-1.D SUBJECT_SAMPLE_FACTORS - wt2 pbs2-2 experimental factors:ctl1 | Sample source:wt2-ripk3-ko-littermate RAW_FILE_NAME(raw data file names)=wt2 pbs2-2.D SUBJECT_SAMPLE_FACTORS - wt2 pbs3-1 experimental factors:ctl1 | Sample source:wt2-ripk3-ko-littermate RAW_FILE_NAME(raw data file names)=wt2 pbs3-1.D SUBJECT_SAMPLE_FACTORS - wt2 pbs3-2 experimental factors:ctl1 | Sample source:wt2-ripk3-ko-littermate RAW_FILE_NAME(raw data file names)=wt2 pbs3-2.D SUBJECT_SAMPLE_FACTORS - wt2 lps1-1 experimental factors:lps | Sample source:wt2-ripk3-ko-littermate RAW_FILE_NAME(raw data file names)=wt2 lps1-1.D SUBJECT_SAMPLE_FACTORS - wt2 lps1-2 experimental factors:lps | Sample source:wt2-ripk3-ko-littermate RAW_FILE_NAME(raw data file names)=wt2 lps1-2.D SUBJECT_SAMPLE_FACTORS - wt2 lps2-1 experimental factors:lps | Sample source:wt2-ripk3-ko-littermate RAW_FILE_NAME(raw data file names)=wt2 lps2-1.D SUBJECT_SAMPLE_FACTORS - wt2 lps2-2 experimental factors:lps | Sample source:wt2-ripk3-ko-littermate RAW_FILE_NAME(raw data file names)=wt2 lps2-2.D SUBJECT_SAMPLE_FACTORS - wt2 lps3-1 experimental factors:lps | Sample source:wt2-ripk3-ko-littermate RAW_FILE_NAME(raw data file names)=wt2 lps3-1.D SUBJECT_SAMPLE_FACTORS - wt2 lps3-2 experimental factors:lps | Sample source:wt2-ripk3-ko-littermate RAW_FILE_NAME(raw data file names)=wt2 lps3-2.D SUBJECT_SAMPLE_FACTORS - Ripk3-ko pbs1-1 experimental factors:ctl1 | Sample source:ripk3-ko RAW_FILE_NAME(raw data file names)=Ripk3-ko pbs1-1.D SUBJECT_SAMPLE_FACTORS - Ripk3-ko pbs1-2 experimental factors:ctl1 | Sample source:ripk3-ko RAW_FILE_NAME(raw data file names)=Ripk3-ko pbs1-2.D SUBJECT_SAMPLE_FACTORS - Ripk3-ko pbs2-1 experimental factors:ctl1 | Sample source:ripk3-ko RAW_FILE_NAME(raw data file names)=Ripk3-ko pbs2-1.D SUBJECT_SAMPLE_FACTORS - Ripk3-ko pbs2-2 experimental factors:ctl1 | Sample source:ripk3-ko RAW_FILE_NAME(raw data file names)=Ripk3-ko pbs2-2.D SUBJECT_SAMPLE_FACTORS - Ripk3-ko pbs3-1 experimental factors:ctl1 | Sample source:ripk3-ko RAW_FILE_NAME(raw data file names)=Ripk3-ko pbs3-1.D SUBJECT_SAMPLE_FACTORS - Ripk3-ko pbs3-2 experimental factors:ctl1 | Sample source:ripk3-ko RAW_FILE_NAME(raw data file names)=Ripk3-ko pbs3-2.D SUBJECT_SAMPLE_FACTORS - Ripk3-ko lps1-1 experimental factors:lps | Sample source:ripk3-ko RAW_FILE_NAME(raw data file names)=Ripk3-ko lps1-1.D SUBJECT_SAMPLE_FACTORS - Ripk3-ko lps1-2 experimental factors:lps | Sample source:ripk3-ko RAW_FILE_NAME(raw data file names)=Ripk3-ko lps1-2.D SUBJECT_SAMPLE_FACTORS - Ripk3-ko lps2-1 experimental factors:lps | Sample source:ripk3-ko RAW_FILE_NAME(raw data file names)=Ripk3-ko lps2-1.D SUBJECT_SAMPLE_FACTORS - Ripk3-ko lps2-2 experimental factors:lps | Sample source:ripk3-ko RAW_FILE_NAME(raw data file names)=Ripk3-ko lps2-2.D SUBJECT_SAMPLE_FACTORS - Ripk3-ko lps3-1 experimental factors:lps | Sample source:ripk3-ko RAW_FILE_NAME(raw data file names)=Ripk3-ko lps3-1.D SUBJECT_SAMPLE_FACTORS - Ripk3-ko lps3-2 experimental factors:lps | Sample source:ripk3-ko RAW_FILE_NAME(raw data file names)=Ripk3-ko lps3-2.D #COLLECTION CO:COLLECTION_SUMMARY BMDMs were isolated from the bone marrow of mouse (Ripk3-ko, mlkl-ko and the CO:COLLECTION_SUMMARY littermates) femurs and tibias and cultured in DMEM medium (Gibco, C11995500BT, CO:COLLECTION_SUMMARY USA) containing 25 ng/mL M-CSF (meilunbio, MA0608, China) and 10% FBS (Gibco, CO:COLLECTION_SUMMARY A5669701, USA) for 7 days, with a medium change at 3.5 days. After successful CO:COLLECTION_SUMMARY differentiation, cells were incubated with PBS (meilunbio, MA0015, China) or 100 CO:COLLECTION_SUMMARY ng/mL LPS (sigma, L2630, USA) for 24 hours. CO:SAMPLE_TYPE Macrophages CO:COLLECTION_LOCATION bone marrow CO:COLLECTION_FREQUENCY once per mice CO:STORAGE_CONDITIONS -80℃ #TREATMENT TR:TREATMENT_SUMMARY After successful differentiation, cells were incubated with PBS (meilunbio, TR:TREATMENT_SUMMARY MA0015, China) or 100 ng/mL LPS (sigma, L2630, USA) for 24 hours. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Post-confluent cells in 6-well plates were homogenized in 0.5 mL chilled 80% SP:SAMPLEPREP_SUMMARY (v/v) methanol containing heptadecanoic acid (Sigma, H3500, USA) as an internal SP:SAMPLEPREP_SUMMARY standard to monitor batch reproducibility. The samples were centrifuged at 12, SP:SAMPLEPREP_SUMMARY 000 rpm for 10 min and the supernatant was transferred to a high recovery glass SP:SAMPLEPREP_SUMMARY sampling vial (CNW, VAAP-31509–1232-100) to vacuum dry at room temperature. SP:SAMPLEPREP_SUMMARY The residue was oximated with 30 μL pyridine containing 20 mg/ml methoxyamine SP:SAMPLEPREP_SUMMARY hydrochloride (Sigma-Aldrich, 226904) at 37°C overnight and further derivatized SP:SAMPLEPREP_SUMMARY with 20 μL N-tert-Butyldimethylsilyl-N-methyltrifluoroacetamide (Sigma-Aldrich, SP:SAMPLEPREP_SUMMARY 394882) at 70°C for 30 min. SP:PROCESSING_STORAGE_CONDITIONS -20℃ SP:EXTRACT_STORAGE -20℃ #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY 1 μL aliquot of the derivatized sample was injected into Agilent 7890A gas CH:CHROMATOGRAPHY_SUMMARY chromatography coupled with Agilent 5975C mass spectrometer. Separation was CH:CHROMATOGRAPHY_SUMMARY achieved on a HP-5ms fused-silica capillary column (30 m × 250 μm i.d.; 5% CH:CHROMATOGRAPHY_SUMMARY diphenyl - 95% Methylpolysiloxane bonded and crosslinked) with helium as the CH:CHROMATOGRAPHY_SUMMARY carrier gas at a constant flow rate of 1 mL/min through the column. CH:CHROMATOGRAPHY_TYPE GC CH:INSTRUMENT_NAME Agilent 7890A CH:COLUMN_NAME Agilent HP5-MS (30 m x 0.25 mm, 0.25 μm) CH:SOLVENT_A none CH:SOLVENT_B none CH:FLOW_GRADIENT none CH:FLOW_RATE 1 mL/min CH:COLUMN_TEMPERATURE 280℃ #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Agilent 5975C MS:INSTRUMENT_TYPE Single quadrupole MS:MS_TYPE EI MS:ION_MODE POSITIVE MS:MS_COMMENTS The temperature of front injection, MSD transfer interface and electron impact MS:MS_COMMENTS (EI) ion source were set to 280°C, 290°C and 230°C, respectively. The GC oven MS:MS_COMMENTS temperature was set to 100°C for 3 min, with an increment rate of 10°C/ min to MS:MS_COMMENTS 140°C, 8°C/min to 260°C, 10°C/min to 310°C and a final 5 min maintenance at MS:MS_COMMENTS 310°C. The electron impact ionization was 70 eV. After 5 min of solvent delay, MS:MS_COMMENTS the mass data was collected at full scan mode (m/z 50–600) and analyzed with MS:MS_COMMENTS software ChemStation. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS peak area MS_METABOLITE_DATA_START Samples wt1 pbs1-1 wt1 pbs1-2 wt1 pbs2-1 wt1 pbs2-2 wt1 pbs3-1 wt1 pbs3-2 wt1 lps1-1 wt1 lps1-2 wt1 lps2-1 wt1 lps2-2 wt1 lps3-1 wt1 lps3-2 mlkl-ko pbs1-1 mlkl-ko pbs1-2 mlkl-ko pbs2-1 mlkl-ko pbs2-2 mlkl-ko pbs3-1 mlkl-ko pbs3-2 mlkl-ko lps1-1 mlkl-ko lps1-2 mlkl-ko lps2-1 mlkl-ko lps2-2 mlkl-ko lps3-1 mlkl-ko lps3-2 wt2 pbs1-1 wt2 pbs1-2 wt2 pbs2-1 wt2 pbs2-2 wt2 pbs3-1 wt2 pbs3-2 wt2 lps1-1 wt2 lps1-2 wt2 lps2-1 wt2 lps2-2 wt2 lps3-1 wt2 lps3-2 Ripk3-ko pbs1-1 Ripk3-ko pbs1-2 Ripk3-ko pbs2-1 Ripk3-ko pbs2-2 Ripk3-ko pbs3-1 Ripk3-ko pbs3-2 Ripk3-ko lps1-1 Ripk3-ko lps1-2 Ripk3-ko lps2-1 Ripk3-ko lps2-2 Ripk3-ko lps3-1 Ripk3-ko lps3-2 Factors experimental factors:ctl1 | Sample source:wt1-mlkl-ko-littermate experimental factors:ctl1 | Sample source:wt1-mlkl-ko-littermate experimental factors:ctl1 | Sample source:wt1-mlkl-ko-littermate experimental factors:ctl1 | Sample source:wt1-mlkl-ko-littermate experimental factors:ctl1 | Sample source:wt1-mlkl-ko-littermate experimental factors:ctl1 | Sample source:wt1-mlkl-ko-littermate experimental factors:lps | Sample source:wt1-mlkl-ko-littermate experimental factors:lps | Sample source:wt1-mlkl-ko-littermate experimental factors:lps | Sample source:wt1-mlkl-ko-littermate experimental factors:lps | Sample source:wt1-mlkl-ko-littermate experimental factors:lps | Sample source:wt1-mlkl-ko-littermate experimental factors:lps | Sample source:wt1-mlkl-ko-littermate experimental factors:ctl1 | Sample source:mlkl-ko experimental factors:ctl1 | Sample source:mlkl-ko experimental factors:ctl1 | Sample source:mlkl-ko experimental factors:ctl1 | Sample source:mlkl-ko experimental factors:ctl1 | Sample source:mlkl-ko experimental factors:ctl1 | Sample source:mlkl-ko experimental factors:lps | Sample source:mlkl-ko experimental factors:lps | Sample source:mlkl-ko experimental factors:lps | Sample source:mlkl-ko experimental factors:lps | Sample source:mlkl-ko experimental factors:lps | Sample source:mlkl-ko experimental factors:lps | Sample source:mlkl-ko experimental factors:ctl1 | Sample source:wt2-ripk3-ko-littermate experimental factors:ctl1 | Sample source:wt2-ripk3-ko-littermate experimental factors:ctl1 | Sample source:wt2-ripk3-ko-littermate experimental factors:ctl1 | Sample source:wt2-ripk3-ko-littermate experimental factors:ctl1 | Sample source:wt2-ripk3-ko-littermate experimental factors:ctl1 | Sample source:wt2-ripk3-ko-littermate experimental factors:lps | Sample source:wt2-ripk3-ko-littermate experimental factors:lps | Sample source:wt2-ripk3-ko-littermate experimental factors:lps | Sample source:wt2-ripk3-ko-littermate experimental factors:lps | Sample source:wt2-ripk3-ko-littermate experimental factors:lps | Sample source:wt2-ripk3-ko-littermate experimental factors:lps | Sample source:wt2-ripk3-ko-littermate experimental factors:ctl1 | Sample source:ripk3-ko experimental factors:ctl1 | Sample source:ripk3-ko experimental factors:ctl1 | Sample source:ripk3-ko experimental factors:ctl1 | Sample source:ripk3-ko experimental factors:ctl1 | Sample source:ripk3-ko experimental factors:ctl1 | Sample source:ripk3-ko experimental factors:lps | Sample source:ripk3-ko experimental factors:lps | Sample source:ripk3-ko experimental factors:lps | Sample source:ripk3-ko experimental factors:lps | Sample source:ripk3-ko experimental factors:lps | Sample source:ripk3-ko experimental factors:lps | Sample source:ripk3-ko L-Phenylalanine 0.042594271 0.041217817 0.030803947 0.031634068 0.036374017 0.036270537 0.109845069 0.13380661 0.273601774 0.312060642 0.114709227 0.123139191 0.118804583 0.119457743 0.101391599 0.117310658 0.116940973 0.130151195 0.043145872 0.042909325 0.039917979 0.04036759 0.02505027 0.021579764 0.061677943 0.065899056 0.05154785 0.04764903 0.127167622 0.122037258 0.096776814 0.092389477 0.09754978 0.093146673 0.115077267 0.122999425 0.094880384 0.085142623 0.074299564 0.075771106 0.113112282 0.0854009 0.077975985 0.079089687 0.057684539 0.059690955 0.140794028 0.163924972 proline 0.171783331 0.173111128 0.189648629 0.185533793 0.227218558 0.224530267 1.583510682 1.489972513 1.365712209 1.362041076 1.574117565 1.582545156 0.847343846 0.908835455 0.841781677 0.925214889 1.040151146 1.142181881 0.18101429 0.188515179 0.195116042 0.175380892 0.143604193 0.151207911 0.66444043 0.661283452 0.797738274 0.768557761 0.695995108 0.793804482 0.942695945 1.191227396 1.109382234 0.864252835 1.37014369 1.238754395 0.438549772 0.44140757 0.484157236 0.488393641 0.75175919 0.669017207 0.515592908 0.572388463 0.507004778 0.550462878 0.70222618 0.606662633 pyruvate 0.223196877 0.223207361 0.183706774 0.178903924 0.187178165 0.209485932 0.791786501 0.985448296 1.712263508 1.357604127 0.86846119 0.975601072 0.503963169 0.397615672 0.376001377 0.54581482 0.348161715 0.504370434 0.285204323 0.294039863 0.291483334 0.319882266 0.184518484 0.209834018 0.247274153 0.315897467 0.275522908 0.286504933 0.512307756 0.318879898 0.522834338 0.520140002 0.459223739 0.576048969 0.556807476 0.600973234 0.219661677 0.218464258 0.177810386 0.172126937 0.192098812 0.212390666 0.196434094 0.244281438 0.226619046 0.227526919 0.324666088 0.329691952 Cholesterol 6.178737231 6.126711854 8.346703669 8.709332205 9.419339982 9.627450685 0.338366411 0.404940011 0.43038785 0.428043164 0.479132043 0.435129641 23.26807602 20.32357043 23.72117741 26.56817951 24.95045998 30.07146219 9.440242395 9.661749963 10.68271212 9.85917098 7.126050114 6.911223521 15.31333754 15.15387505 19.48032999 18.32600584 19.87952161 18.11686467 0.278252584 0.320043291 0.286903198 0.279541961 0.279894435 0.281708109 22.66966474 21.31581787 22.67234036 22.91598669 26.56618528 24.93675986 20.20423365 21.58152209 22.04863389 22.652452 25.18675393 22.08467418 L-Tyrosine 0.338523342 0.331789936 0.268994182 0.264092855 0.322059076 0.321723425 1.417072622 1.45915861 2.550814753 2.522834324 1.332345369 1.408022412 1.024924059 0.875319559 0.956092907 0.983974619 0.942325713 1.072597632 0.314323018 0.313005824 0.321283564 0.300537304 0.171412736 0.171823113 0.65514977 0.706780007 0.557534374 0.509243638 1.285608096 1.118048503 1.053168998 0.986034466 1.195242064 1.014920301 1.088685272 1.305922877 0.724915619 0.723557487 0.660275764 0.773149972 0.985888867 0.842431178 0.634933765 0.685403529 0.54631574 0.521036564 1.26100451 1.14446172 citrate 0.075354067 0.074572453 0.096706255 0.096134416 0.105800342 0.106236544 4.177727743 4.427826708 5.88346969 6.018839406 4.528438863 4.810852142 0.365550394 0.34869163 0.355975267 0.420820738 0.398338149 0.459607567 0.067253771 0.068200697 0.062751116 0.058209942 0.045733716 0.043352348 0.379513966 0.398192391 0.387530058 0.394344884 0.437968033 0.4252236 3.751268642 3.507241262 3.864802159 3.695579978 3.51817558 3.971187565 0.638804163 0.581304574 0.646548905 0.744393081 0.895038127 0.764676074 0.690208113 0.722723871 0.744215712 0.796426538 0.831675951 0.828416715 glutamic acid 1.427497642 1.359201292 1.688668747 1.707053007 2.09574653 2.060576364 4.633868673 6.19936165 5.547515244 5.438935513 5.651571603 5.978150794 4.146620924 3.521158762 3.452849908 4.046896874 4.253401497 4.812529795 1.23691652 1.265803605 1.316675769 1.335470943 0.820590528 0.711790753 7.235346546 7.244272255 8.944900696 8.140069787 8.739250466 8.827719927 4.547481022 5.392662089 4.836740631 4.742845025 5.729959672 5.744487372 3.961493434 3.535479296 4.406688325 4.349536125 4.958851387 4.488456602 4.109444281 4.051372815 4.11482154 4.263582924 4.707860044 4.707908765 malate 0.070227962 0.069584626 0.089465993 0.090135564 0.10320173 0.103007158 0.122781215 0.129689811 0.137769455 0.152694094 0.115249259 0.123178101 0.148469018 0.142495509 0.136695168 0.159398311 0.156864441 0.174623917 0.049577697 0.048638942 0.057445527 0.05848188 0.041434973 0.03980197 0.355250317 0.371115251 0.444473087 0.382143529 0.423024268 0.41637538 0.166012646 0.168791897 0.167097135 0.161996231 0.190859259 0.194656615 0.22559794 0.205193875 0.240292206 0.238260856 0.315344324 0.255779272 0.4684006 0.487880279 0.503092106 0.491766269 0.498243669 0.557779327 ita internal 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 L-Aspartic acid 0.482615795 0.49488679 0.602786689 0.598806299 0.699291668 0.716578419 3.061878253 3.12155467 4.524460133 3.763173405 2.988828188 3.13852489 2.054409025 1.76688744 1.510416188 2.116465303 2.546597422 2.880530818 0.353179099 0.367426211 0.378845308 0.369776297 0.232251983 0.216099987 2.611798656 2.715796665 2.948439972 2.858515327 2.982999864 2.945892546 2.142039986 2.635032465 2.393985662 2.562201685 2.430023214 2.879306867 1.154322863 1.384678693 1.287435428 1.30001357 2.030954059 1.472566013 0.940435502 0.992776261 0.81911586 0.855316551 1.07326342 1.088950247 succinate 0.099974838 0.090299873 0.095702071 0.093852602 0.099050602 0.101710579 0.873676581 0.991274856 1.307079157 1.273746509 1.089745683 1.096792547 0.096350829 0.093427693 0.064818375 0.077402328 0.08905812 0.09937996 0.549777976 0.578048344 0.682542231 0.663382795 0.424836244 0.426045371 0.123733647 0.132289983 0.153540697 0.138818984 0.195490581 0.204447799 0.749290821 0.924368421 1.171455008 0.972885071 0.973131863 0.994937962 0.078607353 0.069093588 0.067611528 0.073081099 0.109881718 0.096925708 0.22549478 0.24311652 0.266957009 0.276488571 0.31767246 0.318363408 fumarate 0.053355714 0.050853824 0.060394807 0.06047251 0.06841399 0.066081296 0.110779852 0.109262319 0.121503887 0.122250969 0.116722833 0.115803508 0.09576644 0.09561061 0.109777057 0.107542589 0.117179461 0.115091894 0.039963816 0.040530817 0.04544172 0.045971802 0.033072417 0.033403927 0.225250942 0.223779587 0.255353569 0.257357504 0.256126596 0.26151327 0.126541706 0.125266931 0.141940856 0.137423347 0.152745018 0.148491863 0.121804607 0.127720824 0.141618049 0.139039185 0.174907564 0.180579883 0.321748859 0.319537498 0.353299948 0.348311553 0.333358843 0.323040042 isoleucine 0.298976849 0.290855157 0.212529241 0.207804629 0.25627242 0.261742582 0.844377376 1.009237999 2.284546196 1.926986408 1.092653061 1.115398988 0.762926278 0.560768982 0.562947607 0.607870586 0.752438424 0.856132848 0.391456278 0.375434881 0.383533372 0.367541806 0.253873711 0.283015525 0.53406443 0.536695198 0.414302977 0.400565694 0.924952382 0.805904315 0.846040359 0.687867581 0.863939252 0.847558983 1.056555002 1.12423883 0.583582519 0.571215283 0.404634854 0.397520699 0.54142171 0.504469056 0.708247538 0.839865698 0.687661024 0.681683472 1.520416572 1.266339879 leucine 0.345217451 0.336820615 0.255178633 0.253242075 0.300714379 0.300959569 0.932405863 1.100559952 2.712551288 2.168972412 1.212692133 1.225089916 1.126793957 0.932710829 0.763536526 1.070573556 1.12081286 1.270146717 0.393565886 0.41743744 0.397999891 0.366275643 0.29303945 0.284407144 0.501236672 0.504713805 0.394427281 0.367205513 1.174330227 0.840163759 0.845361487 0.814718915 0.932609911 0.837190184 1.055535787 0.891142503 0.746776979 0.754855752 0.548332358 0.631949004 0.705719351 0.700222799 0.626530667 0.814430153 0.590012919 0.594735359 1.230932757 1.241119298 valine 0.11220734 0.115448508 0.086722843 0.084844121 0.101915777 0.100094881 0.3857493 0.563466516 0.764043242 0.896638344 0.408708878 0.532215116 0.277627231 0.293209209 0.224502898 0.321975999 0.264727323 0.30079839 0.155517789 0.150118202 0.149316316 0.136028151 0.101953468 0.096085242 0.207057039 0.213128948 0.176080152 0.164339669 0.419512423 0.303168701 0.327303497 0.36277265 0.382670688 0.357903604 0.452982242 0.416137817 0.249294507 0.205457093 0.181316805 0.221776441 0.240643137 0.239204314 0.279282189 0.294530221 0.272470119 0.275287879 0.549929422 0.544107105 glycine 0.341458925 0.317870259 0.30214881 0.309743779 0.420216768 0.422241453 3.56201068 3.511714471 3.704390603 4.4653655 3.098050584 3.217352649 1.237140467 1.130774124 0.922777746 1.083632589 1.065549128 1.205676157 0.564123216 0.726478618 0.615564963 0.590288239 0.434404136 0.456160769 1.548102956 1.699092728 1.699948443 1.353375704 2.312686009 1.756109455 3.038959362 3.0209353 2.660493669 2.219762734 3.867366477 3.996755885 0.937208659 0.993134583 1.012152651 0.962175733 1.339571515 1.109700144 0.996568403 1.068995579 0.887662356 1.059627685 1.287466199 1.289763641 Glutamine 0.01815323 0.016708267 0.017940874 0.019373179 0.027253932 0.02751863 0.111930881 0.14075807 0.116704891 0.136443502 0.096002093 0.105177339 0.035170155 0.031681501 0.032153822 0.040787498 0.029043541 0.042349571 0.039519387 0.040835251 0.034867968 0.033084273 0.023209185 0.020411338 0.031002573 0.033808423 0.033162019 0.031957016 0.049919786 0.040310935 0.070051085 0.072844168 0.064494034 0.072397965 0.165803341 0.169873705 0.028711661 0.035141391 0.022691397 0.021533195 0.029772544 0.026176785 0.051315064 0.054656207 0.064997122 0.061740041 0.073131232 0.071731105 alanine 0.216703738 0.213717899 0.193899606 0.193964103 0.251406763 0.251443323 3.661047169 3.433769358 3.125525638 3.96488386 3.66292831 3.895468718 1.322960422 1.235683793 0.955740704 1.121091082 1.156814446 1.305023371 0.309189193 0.295755807 0.304380927 0.334685826 0.24844155 0.276310994 1.219629964 1.584654572 1.298810679 1.377950961 1.808316043 1.195528386 3.136829644 2.516335329 2.701090322 2.67705507 2.990050156 3.129490897 0.891837532 0.695040048 0.902835089 0.704288837 1.133041866 1.01615635 0.730538535 1.049834375 0.862804901 0.888659136 0.931744287 0.92940397 lysine 0.03321021 0.028416011 0.02853962 0.028211086 0.032273783 0.030968635 0.442201451 0.437947875 0.59841091 0.573315409 0.383750218 0.402495689 0.228071307 0.190820709 0.172996528 0.231471525 0.212930115 0.244978075 0.106950071 0.106751759 0.107442843 0.101845097 0.074085474 0.069166796 0.087285059 0.088371343 0.103563904 0.102808089 0.196581124 0.174638927 0.221013386 0.261025349 0.21662756 0.251800343 0.295127593 0.302550995 0.134798648 0.145689971 0.141255714 0.141176162 0.148598617 0.12598249 0.211896553 0.222466078 0.196576264 0.192905273 0.310759227 0.342995006 methionine 0.025830405 0.024048752 0.019232559 0.019475209 0.022114152 0.022830844 0.079718269 0.097791418 0.211842998 0.219108111 0.096249782 0.109233549 0.089146821 0.07716525 0.06191395 0.08856354 0.072986852 0.101720911 0.037439344 0.035646332 0.033150325 0.032015708 0.021675906 0.020203331 0.040521027 0.043894942 0.031213422 0.029733438 0.078183903 0.083436869 0.071784633 0.073633584 0.07702916 0.074547268 0.091664972 0.093989827 0.054689761 0.054852997 0.042541858 0.052262904 0.070006198 0.060630033 0.05713204 0.075436931 0.051918378 0.053939853 0.125306314 0.105754266 akg 0.017742575 0.017645733 0.021092006 0.021739756 0.023763921 0.022915272 0.026809568 0.030544985 0.035892027 0.041931186 0.027510902 0.036133672 0.052747817 0.043883628 0.046745524 0.058003983 0.054155066 0.062831742 0.01529895 0.016852199 0.018573499 0.019794834 0.016736139 0.016773983 0.091509739 0.091479778 0.122758267 0.117204373 0.109974745 0.113338987 0.035852918 0.045264157 0.055729034 0.049563997 0.048611394 0.052486301 0.060349388 0.061151371 0.059314639 0.063225762 0.075462426 0.076425609 0.129491793 0.12895594 0.139227647 0.145095423 0.183190245 0.18847232 lactate 0.979178603 0.995551798 0.816054849 0.807266538 0.884068246 0.910773264 5.996494565 8.092620989 9.084831425 9.162789519 6.480929356 6.867445997 2.014502201 1.469537685 1.332120108 1.862973145 1.432283197 1.872032118 1.285580562 1.211012014 1.299922834 1.179261395 1.086371364 0.864947575 1.450125826 1.378595031 1.292042009 1.183683821 1.915171511 1.752763336 5.203260888 4.705453282 5.405786594 5.243900399 5.375044903 6.135261815 1.2349729 1.232201618 1.186196055 1.111539382 1.594325784 1.144538264 2.582662718 3.252360328 3.432257053 3.316575165 3.299275823 3.826422291 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name retention index quantified m/z pubchem identifieres KEGG identifiers L-Phenylalanine 336 6140 C00079  proline 184 145742 C00148   pyruvate 174 107735 C00022 Cholesterol 443 5997 C00187  L-Tyrosine 302 6057 C00082  citrate 459 31348 C00158  glutamic acid 432 33032 C00025    malate 419 160434 C00149      L-Aspartic acid 302 5960 C00049  succinate 289 160419 C00042  fumarate 287 5460307 C00122    isoleucine 200 6306 C00407 leucine 200 6106 C00123 valine 260 6287 C00183  glycine 218 750 C00037 Glutamine 431 5961 C00303  alanine 158 5950 C00041 lysine 300 5962 C00047  methionine 292 6137 C00073 akg 346 51 C00026   lactate 261 91435 C00186  ita internal 306 162342694 none METABOLITES_END #END