#METABOLOMICS WORKBENCH nozomu_kono_20250125_170427 DATATRACK_ID:5571 STUDY_ID:ST003749 ANALYSIS_ID:AN006156 PROJECT_ID:PR002333 VERSION 1 CREATED_ON February 24, 2025, 5:24 pm #PROJECT PR:PROJECT_TITLE Phosphatidylinositol remodeling by lysophospholipid acyltransferase 11 ensures PR:PROJECT_TITLE embryonic radial glial cell integrity PR:PROJECT_SUMMARY Arachidonic acid, a vital polyunsaturated fatty acid in brain development, is PR:PROJECT_SUMMARY enriched in phosphatidylinositol (PI). The arachidonic acyl chain in PI is PR:PROJECT_SUMMARY introduced by lysophospholipid acyltransferase 11 (LPLAT11)/membrane-bound PR:PROJECT_SUMMARY O-acyltransferase 7 (MBOAT7), the loss of which causes microcephaly in humans PR:PROJECT_SUMMARY and mice. Here, we show that LPLAT11 deficiency impaired indirect neurogenesis PR:PROJECT_SUMMARY in the developing neocortex, resulting in fewer layer II-V neurons. PR:PROJECT_SUMMARY LPLAT11-deficient radial glial cells had defects in differentiation into PR:PROJECT_SUMMARY intermediate progenitor cells and increased apoptosis. Prior to these anomalies, PR:PROJECT_SUMMARY LPLAT11 deficiency caused a rounding of the Golgi apparatus, accompanied by PR:PROJECT_SUMMARY impaired apical trafficking of E-cadherin, and deregulated apical detachment. PR:PROJECT_SUMMARY Moreover, impaired PI acyl chain remodeling led to a decreased amount of PR:PROJECT_SUMMARY PI(4,5)P2, leading to Golgi apparatus rounding. Thus, these results clarify the PR:PROJECT_SUMMARY underlying mechanism of microcephaly by LPLAT11 deficiency and highlight the PR:PROJECT_SUMMARY critical role of arachidonic acid in PI in the integrity of radial glial cells. PR:INSTITUTE University of Tokyo PR:LAST_NAME Nozomu PR:FIRST_NAME Kono PR:ADDRESS Hongo 7-3-1, Bunkyo-ku, Tokyo 113-0033, JAPAN PR:EMAIL nozomu@mol.f.u-tokyo.ac.jp PR:PHONE 81-3-5841-4723 #STUDY ST:STUDY_TITLE Phosphatidylinositol remodeling by lysophospholipid acyltransferase 11 ensures ST:STUDY_TITLE embryonic radial glial cell integrity - part 3 ST:STUDY_SUMMARY To further examine whether PI(4,5)P2 is needed for the proper morphology of the ST:STUDY_SUMMARY Golgi apparatus in RGCs, we treated E12.5 hemispheres with UNC3230, a PIPKIγ ST:STUDY_SUMMARY inhibitor. UNC3230 significantly reduced the total amount of PI(4,5)P2, ST:STUDY_SUMMARY primarily affecting PI(4,5)P2 species containing arachidonic acid and its ST:STUDY_SUMMARY analogous fatty acid, Mead acid (20:3), while exerting minimal effect on other ST:STUDY_SUMMARY PI(4,5)P2 molecular species. UNC3230 treatment caused rounding of the Golgi ST:STUDY_SUMMARY apparatus, as observed in Mboat7 KO mice. We then administered UNC3230 into the ST:STUDY_SUMMARY lateral ventricle at E12.5 and immunostained E13.5 cortices with antibodies ST:STUDY_SUMMARY against PI(4,5)P2, GM130, E-cadherin, Sox2, and p-H3. UNC3230 administration ST:STUDY_SUMMARY caused a decreased level of PI(4,5)P2, a rounding of the Golgi apparatus, and ST:STUDY_SUMMARY reduced apical expression of E-cadherin in the ventricular zone, whereas the ST:STUDY_SUMMARY detachment of RGCs from the ventricular wall was not apparent. These results ST:STUDY_SUMMARY suggest that reduced PI(4,5)P2 levels cause rounding of the Golgi apparatus and ST:STUDY_SUMMARY decreased E-cadherin expression at the apical surface. We also treated E12.5 ST:STUDY_SUMMARY Mboat7 heterozygous hemispheres with Sevenin-1, an LPLAT11 inhibitor. Sevenin-1 ST:STUDY_SUMMARY treatment caused rounding of the Golgi apparatus in the Mboat7 heterozygous ST:STUDY_SUMMARY hemispheres, whereas it had no effect on the morphology of the Golgi apparatus ST:STUDY_SUMMARY in the Mboat7 KO hemispheres, highlighting the importance of LPLAT11 activity. ST:STUDY_SUMMARY SFC-MS analysis confirmed that Sevenin-1 decreased the amount of PI with ST:STUDY_SUMMARY arachidonic acid and the total amount of PI(4,5)P2. ST:INSTITUTE University of Tokyo ST:LAST_NAME Nozomu ST:FIRST_NAME Kono ST:ADDRESS Hongo 7-3-1, Bunkyo-ku, Tokyo 113-0033, JAPAN ST:EMAIL nozomu@mol.f.u-tokyo.ac.jp ST:PHONE 81-3-5841-4723 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 #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 - DMSO(Sevenin-1 control)_1 Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:DMSO RAW_FILE_NAME(Raw file name)=E12.5 hemisphere culture PIPs DMSO_S 1.mzML SUBJECT_SAMPLE_FACTORS - DMSO(Sevenin-1 control)_2 Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:DMSO RAW_FILE_NAME(Raw file name)=E12.5 hemisphere culture PIPs DMSO_S 2.mzML SUBJECT_SAMPLE_FACTORS - DMSO(Sevenin-1 control)_3 Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:DMSO RAW_FILE_NAME(Raw file name)=E12.5 hemisphere culture PIPs DMSO_S 3.mzML SUBJECT_SAMPLE_FACTORS - DMSO(Sevenin-1 control)_4 Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:DMSO RAW_FILE_NAME(Raw file name)=E12.5 hemisphere culture PIPs DMSO_S 4.mzML SUBJECT_SAMPLE_FACTORS - DMSO(Sevenin-1 control)_5 Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:DMSO RAW_FILE_NAME(Raw file name)=E12.5 hemisphere culture PIPs DMSO_S 5.mzML SUBJECT_SAMPLE_FACTORS - Sevenin-1_1 Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:Sevenin-1 RAW_FILE_NAME(Raw file name)=E12.5 hemisphere culture PIPs Sevenin-1 1.mzML SUBJECT_SAMPLE_FACTORS - Sevenin-1_2 Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:Sevenin-1 RAW_FILE_NAME(Raw file name)=E12.5 hemisphere culture PIPs Sevenin-1 2.mzML SUBJECT_SAMPLE_FACTORS - Sevenin-1_3 Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:Sevenin-1 RAW_FILE_NAME(Raw file name)=E12.5 hemisphere culture PIPs Sevenin-1 3.mzML SUBJECT_SAMPLE_FACTORS - Sevenin-1_4 Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:Sevenin-1 RAW_FILE_NAME(Raw file name)=E12.5 hemisphere culture PIPs Sevenin-1 4.mzML SUBJECT_SAMPLE_FACTORS - Sevenin-1_5 Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:Sevenin-1 RAW_FILE_NAME(Raw file name)=E12.5 hemisphere culture PIPs Sevenin-1 5.mzML SUBJECT_SAMPLE_FACTORS - DMSO (UNC3230 control)_1 Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:DMSO RAW_FILE_NAME(Raw file name)=E12.5 hemisphere culture PIPs DMSO_U 1.mzML SUBJECT_SAMPLE_FACTORS - DMSO (UNC3230 control)_2 Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:DMSO RAW_FILE_NAME(Raw file name)=E12.5 hemisphere culture PIPs DMSO_U 2.mzML SUBJECT_SAMPLE_FACTORS - DMSO (UNC3230 control)_3 Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:DMSO RAW_FILE_NAME(Raw file name)=E12.5 hemisphere culture PIPs DMSO_U 3.mzML SUBJECT_SAMPLE_FACTORS - DMSO (UNC3230 control)_4 Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:DMSO RAW_FILE_NAME(Raw file name)=E12.5 hemisphere culture PIPs DMSO_U 4.mzML SUBJECT_SAMPLE_FACTORS - DMSO (UNC3230 control)_5 Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:DMSO RAW_FILE_NAME(Raw file name)=E12.5 hemisphere culture PIPs DMSO_U 5.mzML SUBJECT_SAMPLE_FACTORS - DMSO (UNC3230 control)_6 Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:DMSO RAW_FILE_NAME(Raw file name)=E12.5 hemisphere culture PIPs DMSO_U 6.mzML SUBJECT_SAMPLE_FACTORS - UNC3230_1 Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:UNC3230 RAW_FILE_NAME(Raw file name)=E12.5 hemisphere culture PIPs UNC3230 1.mzML SUBJECT_SAMPLE_FACTORS - UNC3230_2 Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:UNC3230 RAW_FILE_NAME(Raw file name)=E12.5 hemisphere culture PIPs UNC3230 2.mzML SUBJECT_SAMPLE_FACTORS - UNC3230_3 Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:UNC3230 RAW_FILE_NAME(Raw file name)=E12.5 hemisphere culture PIPs UNC3230 3.mzML SUBJECT_SAMPLE_FACTORS - UNC3230_4 Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:UNC3230 RAW_FILE_NAME(Raw file name)=E12.5 hemisphere culture PIPs UNC3230 4.mzML SUBJECT_SAMPLE_FACTORS - UNC3230_5 Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:UNC3230 RAW_FILE_NAME(Raw file name)=E12.5 hemisphere culture PIPs UNC3230 5.mzML SUBJECT_SAMPLE_FACTORS - UNC3230_6 Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:UNC3230 RAW_FILE_NAME(Raw file name)=E12.5 hemisphere culture PIPs UNC3230 6.mzML #COLLECTION CO:COLLECTION_SUMMARY The brains of embryos were dissected in individual 35 mm dishes containing CO:COLLECTION_SUMMARY culture medium (Opti-MEM I with 20 mM glucose, 55 µM 2-mercaptoethanol, and 1% CO:COLLECTION_SUMMARY penicillin/streptomycin). Separate hemispheres were transferred to a new 35m CO:COLLECTION_SUMMARY dish containing culture medium using a P1000 Pipetman with a cut pipette tip and CO:COLLECTION_SUMMARY kept on ice until every dissection was completed. Hemispheres were transferred CO:COLLECTION_SUMMARY to an individual well of a 12-well plate containing 2 ml of culture medium. CO:COLLECTION_SUMMARY Plates were then placed on a nutator inside a 5% CO2 incubator for 24 h. CO:SAMPLE_TYPE Cultured hemisphere #TREATMENT TR:TREATMENT_SUMMARY N-(2-(Bicyclo[2.2.1]heptanyl)ethyl)-4-(2-(N-methylsulfamoyl)phenyl)piperazine-1-carboxamide TR:TREATMENT_SUMMARY (Sevenin-1) was added at 10 µM final concentration in the culture medium. TR:TREATMENT_SUMMARY UNC3230 was added at 0.25 µM final concentration in the culture medium. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Cultured hemispheres were collected into a safe-lock poly-propylene tube (2ml) SP:SAMPLEPREP_SUMMARY with 750 µl of ice-cold quench mix and sonicated. 170 µl of H2O was mixed and SP:SAMPLEPREP_SUMMARY stored at -80 ºC before lipid extraction. The single-phase sample (a mixture of SP:SAMPLEPREP_SUMMARY 170 µl of an aqueous sample, 10 ng of internal standards [17:0/20:4 PI, SP:SAMPLEPREP_SUMMARY 17:0/20:4 PI4P, and 17:0/20:4 PI(4,5)P2], and 750 µl of quench mix) were mixed SP:SAMPLEPREP_SUMMARY with 725 µl of CHCl3 and 170 µl of 2 M HCl, followed by vortex-mixing and SP:SAMPLEPREP_SUMMARY centrifugation (1,500 g, 5 min at room temperature). The lower organic phase was SP:SAMPLEPREP_SUMMARY collected into a new 2 ml safe-lock poly-propylene tube and mixed with 708 µl SP:SAMPLEPREP_SUMMARY of pre-derivatization wash solution, followed by vortex-mixing and SP:SAMPLEPREP_SUMMARY centrifugation (1,500 g, 3 min at room temperature). The lower phase was SP:SAMPLEPREP_SUMMARY collected into another fresh tube and subjected to derivatization. Fifty µl SP:SAMPLEPREP_SUMMARY trimethylsilyl diazomethane in hexane (2 M solution) was added to the lipid SP:SAMPLEPREP_SUMMARY extracts (~1 ml), and after leaving to stand 10 min at room temperature, the SP:SAMPLEPREP_SUMMARY reaction was quenched with 6 µl of acetic acid. Next, 700 µl SP:SAMPLEPREP_SUMMARY post-derivatization wash solution was added to the mixture, followed by SP:SAMPLEPREP_SUMMARY centrifugation (1,500 g, 3 min). The resultant lower phase was collected and SP:SAMPLEPREP_SUMMARY rewashed with a 700 µl post-derivatization wash solution. Samples were SP:SAMPLEPREP_SUMMARY temporarily stored at -80 ºC. Ninety µl of methanol and 10 µl of H2O were SP:SAMPLEPREP_SUMMARY added to the final collected lower phase. The samples were dried up with a SP:SAMPLEPREP_SUMMARY centrifugal evaporator. Samples were dissolved in 80 µl methanol, sonicated for SP:SAMPLEPREP_SUMMARY 1 min, and 20 µl H2O was added. After centrifugation at 15,000 g for 30 min at SP:SAMPLEPREP_SUMMARY 4 ºC, the supernatants were collected. To avoid degradation, the samples were SP:SAMPLEPREP_SUMMARY stored at -80 ºC until use. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Supercritical fluid chromatography CH:INSTRUMENT_NAME Shimadzu Nexera UC/s CH:COLUMN_NAME ULTRON AF-HILIC-CD (250 x 2.0mm, 5.0um) CH:SOLVENT_A 99.99% supercritical carbon dioxide CH:SOLVENT_B 2.5% water/97.5% methanol; 0.1% formic acid CH:FLOW_GRADIENT 0-16 min: 5% B to 20% B; 16.01-18 min: 40% B; 18.01-22 min: 5% B CH:FLOW_RATE 1.5 mL/min CH:COLUMN_TEMPERATURE 10 ºC #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME ABI Sciex 4500 QTrap MS:INSTRUMENT_TYPE Triple quadrupole MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS The instrument parameters of QTRAP4500 for positive ion mode were as follows: MS:MS_COMMENTS curtain gas, 20 psi; ionspray voltage, 4500 V; temperature, 300 ºC; ion source MS:MS_COMMENTS gas 1, 18 psi; ion source gas 2, 20 psi. The specific detection of PI and MS:MS_COMMENTS phosphoinositide species was performed using MRM. 17:0/20:4 PI, 17:0/20:4 PI4P, MS:MS_COMMENTS and 17:0/20:4 PI4,5P2 ware used as internal standards. Analyst (SCIEX) was used MS:MS_COMMENTS for data acquisition and processing. MultiQuant (SCIEX) was used for data MS:MS_COMMENTS evaluation. Gaussian smoothing width was 2.0 points. Retention time of PI and MS:MS_COMMENTS PI(4,5)P2 is confirmed by measuring standards [17:0/20:4 PI, PI(3,4)P2, MS:MS_COMMENTS PI(3,5)P2, and PI(4,5)P2] before measuring samples. Peak areas of PI and MS:MS_COMMENTS PI(4,5)P2 were integrated from the ion chromatogram of PI and of PIP2, MS:MS_COMMENTS respectively, based on retention time, and isotopic correction was applied. The MS:MS_COMMENTS peak area of the lipid species was divided by the corresponding internal MS:MS_COMMENTS standard area, and the total amount of phosphoinositides per hemisphere was MS:MS_COMMENTS calculated from the ratio of injection volume to total sample volume. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS pmol/hemisphere MS_METABOLITE_DATA_START Samples DMSO(Sevenin-1 control)_1 DMSO(Sevenin-1 control)_2 DMSO(Sevenin-1 control)_3 DMSO(Sevenin-1 control)_4 DMSO(Sevenin-1 control)_5 Sevenin-1_1 Sevenin-1_2 Sevenin-1_3 Sevenin-1_4 Sevenin-1_5 DMSO (UNC3230 control)_1 DMSO (UNC3230 control)_2 DMSO (UNC3230 control)_3 DMSO (UNC3230 control)_4 DMSO (UNC3230 control)_5 DMSO (UNC3230 control)_6 UNC3230_1 UNC3230_2 UNC3230_3 UNC3230_4 UNC3230_5 UNC3230_6 Factors Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:DMSO Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:DMSO Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:DMSO Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:DMSO Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:DMSO Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:Sevenin-1 Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:Sevenin-1 Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:Sevenin-1 Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:Sevenin-1 Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:Sevenin-1 Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:DMSO Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:DMSO Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:DMSO Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:DMSO Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:DMSO Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:DMSO Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:UNC3230 Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:UNC3230 Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:UNC3230 Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:UNC3230 Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:UNC3230 Sample source:Brain hemisphere culture | Gestational ages:E12.5 | Treatment:UNC3230 PI 32:1 10.02387716 9.056899826 16.78706995 15.74147596 7.417416189 7.422089159 9.437889927 17.11279699 14.42281316 11.54910618 ND ND ND ND ND ND ND ND ND ND ND ND PI 34:0 1.192857718 2.0282416 5.160876404 3.242964562 0.660332765 2.504825214 1.823377155 5.441589775 3.388596378 2.050670314 ND ND ND ND ND ND ND ND ND ND ND ND PI 34:1 46.38342736 59.13192924 45.16852569 44.58573343 39.56531885 59.59142746 75.32131097 97.55938038 45.11030203 73.10165877 ND ND ND ND ND ND ND ND ND ND ND ND PI 34:2 27.79510563 39.02808871 62.312485 41.78111039 25.16180592 31.0099847 34.33583129 62.73970337 30.28405579 21.75566888 ND ND ND ND ND ND ND ND ND ND ND ND PI 36:1 23.92801419 45.30187958 31.06422495 45.12971349 21.4371693 42.96803793 55.71754561 52.78759022 42.70081505 29.56384057 ND ND ND ND ND ND ND ND ND ND ND ND PI 36:2 91.63746756 114.8529688 189.4909797 109.7582885 104.5243443 109.8560156 195.4355542 254.3178883 155.2461934 99.41331712 ND ND ND ND ND ND ND ND ND ND ND ND PI 36:3 89.53487841 84.10335759 101.9217651 96.60107907 68.14614124 56.72564637 91.83524379 105.4692655 61.68952784 105.8734778 ND ND ND ND ND ND ND ND ND ND ND ND PI 36:4 93.35222077 104.896035 164.6574736 96.67110598 89.23645517 82.06028942 119.7295807 186.524122 98.30900562 97.81775351 ND ND ND ND ND ND ND ND ND ND ND ND PI 38:2 43.43753089 45.70415119 45.27072886 31.9164218 36.77181187 31.11125096 45.8891367 34.36467053 27.84838966 47.685816 ND ND ND ND ND ND ND ND ND ND ND ND PI 38:3 190.518301 328.0105166 340.5008048 361.8737281 248.6531803 198.3735782 379.2445579 305.0487676 169.1843659 193.9264167 ND ND ND ND ND ND ND ND ND ND ND ND PI 38:4 507.0444209 784.5470304 862.0873745 560.9003915 635.3239379 494.2772012 711.1674204 814.2630911 562.4802948 736.9075682 ND ND ND ND ND ND ND ND ND ND ND ND PI 38:5 229.5902866 336.9693227 382.4142575 238.5217354 280.6323133 216.976419 334.2176028 376.1751607 298.8476045 282.0179299 ND ND ND ND ND ND ND ND ND ND ND ND PI 38:6 27.11657442 34.04180492 42.73799038 31.65428638 30.01363397 29.52608566 39.02252269 59.64875363 32.70478377 47.67773505 ND ND ND ND ND ND ND ND ND ND ND ND PI 40:3 8.652343225 11.19853594 13.75064756 8.988869806 10.43987636 8.918315773 9.969302351 15.51119356 9.023725695 9.525202871 ND ND ND ND ND ND ND ND ND ND ND ND PI 40:4 10.57761022 17.18089036 20.90289461 15.63852767 14.30182939 15.07848798 20.09540889 22.54160422 13.64570952 15.12382465 ND ND ND ND ND ND ND ND ND ND ND ND PI 40:5 34.30900522 47.79360201 48.22462428 40.98497643 31.51270376 30.02850817 43.94188671 45.66275406 32.05592885 28.30005708 ND ND ND ND ND ND ND ND ND ND ND ND PI 40:6 36.33226191 52.24820713 48.05961273 42.32248149 33.51146349 30.22714033 63.15399754 59.07879126 35.21285762 39.48251019 ND ND ND ND ND ND ND ND ND ND ND ND PI4,5P2 32:0 ND ND ND ND ND ND ND ND ND ND 0.228092855 0.358502807 0.46668084 0.471766445 0.436039244 0.240134998 0.326006539 0.747278851 1.12022975 0.647891644 0.682301048 0.324138064 PI4,5P2 32:1 2.563609071 3.634447119 4.555182464 4.219382281 2.282838834 3.044884973 3.197788488 4.178438285 3.374492549 2.80378924 2.503539373 1.913087909 2.257323043 1.956734926 3.033412084 2.03094859 2.281744457 3.388314106 4.385606491 2.030940301 2.225661417 1.283368663 PI4,5P2 32:2 ND ND ND ND ND ND ND ND ND ND 0.461056467 0.416844649 0.422751108 0.556977174 0.794735529 0.655762509 0.465523899 0.512659285 0.468189573 0.335167242 0.31218599 0.246517684 PI4,5P2 34:0 0.033494249 1.152266664 0.024669202 0.596864676 0.136611185 0.464829115 1.126844523 0 0 0.189565705 0 0.111266639 0 0 0.315245475 0 0 0.40645142 1.159806432 0.739500882 0.407271617 0.195818193 PI4,5P2 34:1 22.69826679 30.65575905 33.95768025 21.24960405 17.32451332 25.38306787 29.16902405 29.85348904 32.49909488 18.98447607 17.09047584 15.14601668 15.44759724 23.26573458 26.44601425 19.33302497 15.52074751 20.81611949 20.15869808 16.78403165 16.62631445 8.211888339 PI4,5P2 34:2 8.623648807 11.68148256 11.60181366 9.24767592 8.349497254 8.740000462 9.576810974 10.48969194 10.27620998 8.797098048 6.975155791 6.803518427 7.438014243 11.26610128 10.26149238 7.200481765 6.206360116 6.007773911 6.161284088 6.098385051 6.535448104 5.486881329 PI4,5P2 36:1 19.31700422 32.06797513 20.21822993 23.23605887 14.3673637 15.09265833 28.80923292 19.62244009 17.51149905 17.59755411 21.24491947 13.80866546 16.17926023 19.44416261 23.17263018 12.06777652 21.27106411 22.98173909 20.10447104 18.15705201 19.28792487 13.39700218 PI4,5P2 36:2 46.39576861 66.49651031 69.73238325 70.21233493 43.31498786 50.62889432 66.85946458 59.58135842 57.65795057 54.37201811 41.69198144 35.14330344 37.64695499 57.53953587 49.2133547 29.16690528 41.54396541 48.82494207 52.90183023 44.94000093 44.47049914 36.76247649 PI4,5P2 36:3 21.8189041 33.24709915 33.78622348 29.2307097 27.09909436 14.74853205 17.83589696 18.5810068 16.12477142 17.72455162 18.45786069 20.26016091 18.09453655 22.27630839 23.81264717 19.96623846 20.62946045 18.92747819 20.67518311 17.35533684 17.56647593 13.34452619 PI4,5P2 36:4 19.96042715 39.65193963 42.35693905 26.34064345 35.94076732 18.04893458 26.9084592 28.47466251 18.73405519 27.96092262 27.75845585 17.81243879 14.40548191 19.13112973 32.44175934 26.62018678 19.02018802 16.45758624 26.25735946 16.11327609 16.18268912 18.46193691 PI4,5P2 36:5 ND ND ND ND ND ND ND ND ND ND 0.915350626 0.647720323 0.573864368 1.011970881 0.745476151 0.627840451 0.759242845 0.510717006 0.820068613 0.424284976 0.453188007 0.541159098 PI4,5P2 38:2 23.15259546 38.00497576 32.77113826 21.79095295 21.5486332 13.58572322 21.36324768 17.67309844 16.69801514 15.53175409 26.51746848 31.391312 17.80398973 25.84040327 36.63933638 25.01276751 16.64577392 25.51655893 21.71514378 26.00629269 21.38802063 15.48091336 PI4,5P2 38:3 80.6580729 129.1851638 109.8719774 91.76005155 86.49644182 50.93343625 77.01488777 69.7115623 63.64784364 80.80787463 93.88750193 88.21214065 81.90068745 122.7616447 138.9293764 91.37797196 55.80678314 67.66754947 66.63583295 49.78800137 62.24078186 51.83055102 PI4,5P2 38:4 204.7899007 273.4353094 257.7810762 199.7876159 254.2529477 152.8491965 211.6284217 192.1616249 161.4009913 196.2889232 199.9949635 209.1216337 282.6373813 218.6919228 299.8576544 189.7587155 165.6451632 201.9819499 140.0856337 133.1484109 203.8707859 183.0077034 PI4,5P2 38:5 42.61167851 60.53431491 60.10708539 48.43954275 64.0186273 35.89358405 49.41055812 42.3796051 33.04343508 54.60741276 30.30806101 26.98241793 56.22346412 34.74418701 51.45751684 33.45914514 38.30979796 38.23076111 33.88012377 28.15042922 43.06908129 36.16413608 PI4,5P2 38:6 1.369819985 1.917167912 2.1517065 1.5933337 2.047817428 1.420666742 1.896352046 1.796180127 1.408396844 2.059332503 1.183958421 0.719734711 1.289778685 0.964788169 1.213440243 1.22483677 1.065340751 0.78897395 1.066406613 0.616587385 0.714275673 0.933755586 PI4,5P2 40:3 1.779433191 2.552214562 2.077033342 1.997827236 1.828683694 1.450561223 1.466212869 1.251232489 1.300814117 1.175191245 1.480384593 1.373762146 1.550228541 2.303726319 1.723745534 1.552483003 1.850692091 1.846080754 1.593318417 1.800041213 1.438108479 1.388029958 PI4,5P2 40:4 3.51707245 5.613446474 5.685334674 3.987399189 5.320093732 2.870678043 3.585221785 3.259804581 3.342570268 4.257486911 3.889685693 3.678828301 2.594009163 3.916924263 4.868794602 3.928181399 3.997726175 3.511097318 5.023893366 3.412836001 3.269150643 2.937603061 PI4,5P2 40:5 2.73309458 4.684411748 4.090717883 3.358122145 3.958426301 2.234293545 3.340344119 2.934739069 2.248405388 3.347978264 4.395189782 2.860941626 1.618678107 3.077275412 4.089913648 2.692718152 2.60618453 2.4170979 3.207911224 1.844908224 1.975822494 2.388184113 PI4,5P2 40:6 2.057055779 3.474442882 2.925663025 2.357382734 2.616170404 2.0377799 2.747586677 2.674849193 1.921923683 2.450914375 1.956393734 1.674791347 1.263356533 2.85508392 2.124411436 1.130761242 1.639659316 1.524631783 1.712465708 1.668836748 1.418218188 1.331014061 PI4,5P2 40:7 ND ND ND ND ND ND ND ND ND ND 0.408177426 0.229034397 0.26004727 0.358416593 0.446232014 0.222645942 0.241952389 0.198623527 0.314922672 0.295893643 0.272096281 0.154306409 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name PI 32:1 PI 34:0 PI 34:1 PI 34:2 PI 36:1 PI 36:2 PI 36:3 PI 36:4 PI 38:2 PI 38:3 PI 38:4 PI 38:5 PI 38:6 PI 40:3 PI 40:4 PI 40:5 PI 40:6 PI4,5P2 32:0 PI4,5P2 32:1 PI4,5P2 32:2 PI4,5P2 34:0 PI4,5P2 34:1 PI4,5P2 34:2 PI4,5P2 36:1 PI4,5P2 36:2 PI4,5P2 36:3 PI4,5P2 36:4 PI4,5P2 36:5 PI4,5P2 38:2 PI4,5P2 38:3 PI4,5P2 38:4 PI4,5P2 38:5 PI4,5P2 38:6 PI4,5P2 40:3 PI4,5P2 40:4 PI4,5P2 40:5 PI4,5P2 40:6 PI4,5P2 40:7 METABOLITES_END #END