#METABOLOMICS WORKBENCH JulieJacquemyn_20210622_094602 DATATRACK_ID:2712 STUDY_ID:ST001859 ANALYSIS_ID:AN003014 PROJECT_ID:PR001172 VERSION 1 CREATED_ON July 16, 2021, 11:35 am #PROJECT PR:PROJECT_TITLE Torsin and NEP1R1-CTDNEP1 affect interphase NPC insertion by lipid-dependent and PR:PROJECT_TITLE -independent mechanisms PR:PROJECT_TYPE MS analysis performed by Lipotype on Drosophila larval fat body tissue PR:PROJECT_SUMMARY The interphase nuclear envelope is extensively remodeled during nuclear pore PR:PROJECT_SUMMARY complex insertion. The process is relatively poorly understood, including why it PR:PROJECT_SUMMARY requires the Torsin ATPases that also regulate NE-localized lipid metabolism. PR:PROJECT_SUMMARY Here we show that fly dTorsin affects lipid metabolism through the PR:PROJECT_SUMMARY NEP1R1-CTDNEP1 phosphatase/ Lipin phosphatidic acid phosphatase pathway. At a PR:PROJECT_SUMMARY cellular level, fly and mouse Torsins removed NEP1R1-CTDNEP1 from the NE to, in PR:PROJECT_SUMMARY turn, exclude Lipin from the nucleus. NEP1R1-CTDNEP1 downregulation also PR:PROJECT_SUMMARY restored nuclear pore membrane fusion in post-mitotic dTorsinKO fat body cells. PR:PROJECT_SUMMARY However, Lipin downregulation was ineffective and membrane fusion defects did PR:PROJECT_SUMMARY not correlate with lipidomic abnormalities. Further testing confirmed that PR:PROJECT_SUMMARY membrane fusion continued in cells with excess Lipin function. It also led to PR:PROJECT_SUMMARY the surprising finding that excess PA metabolism inhibited recruitment of the PR:PROJECT_SUMMARY inner ring complex Nup35 subunit, resulting in elongated channel-like structures PR:PROJECT_SUMMARY in place of mature nuclear pores. We conclude that the NEP1R1-CTDNEP1 PR:PROJECT_SUMMARY phosphatase affects interphase NPC biogenesis by lipid-dependent and PR:PROJECT_SUMMARY lipid-independent mechanisms and this explains some of the pleiotropic effects PR:PROJECT_SUMMARY of Torsins. PR:INSTITUTE VIB KULeuven PR:DEPARTMENT Dept. of Neurosciences, KU Leuven, 3000 Leuven, Belgium PR:LAST_NAME Jacquemyn PR:FIRST_NAME Julie PR:ADDRESS ON 4, 6e verd Campus Gasthuisberg, Herestraat 49, bus 602, 3000 Leuven, Belgium PR:EMAIL julie.jacquemyn@kuleuven.be PR:PHONE 032479570951 #STUDY ST:STUDY_TITLE dTor affects the fat body lipidome via Nep1r1, Ctdnep1 and Lipin ST:STUDY_SUMMARY Quantitative MS analysis was performed on ten 4 day-old Drosophila larval fat ST:STUDY_SUMMARY bodies homogenized in 50µl D-PBS (Dulbecco’s Phosphate Buffered Saline ST:STUDY_SUMMARY without Mg2+ and Ca2+) by Lipotype using previously described methods (Grillet ST:STUDY_SUMMARY et al, 2016). ST:INSTITUTE VIB KULeuven ST:DEPARTMENT Dept. of Neurosciences, KU Leuven, 3000 Leuven, Belgium ST:LAST_NAME Jacquemyn ST:FIRST_NAME Julie ST:ADDRESS ON 4, 6e verd Campus Gasthuisberg, Herestraat 49, bus 602, Leuven, NA, 3000, ST:ADDRESS Belgium ST:EMAIL julie.jacquemyn@kuleuven.be ST:PHONE 0032479570951 #SUBJECT SU:SUBJECT_TYPE Invertebrate SU:SUBJECT_SPECIES Drosophila melanogaster SU:TAXONOMY_ID 7227 SU:AGE_OR_AGE_RANGE Developmentally-staged-larvae were collected (randomized) 4 days after placing SU:AGE_OR_AGE_RANGE male and female flies in fresh vials for a 12-hour period. SU:GENDER Male and female #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 Drosophila s5-1 Genotype:w- Luciferase Sample Description=fat body tissues of 4DO larvae collected and homogenized in D PBS without Mg2+, Ca2+; StartVolume=50 µL; Concentration=10 fat bodies/50µL SUBJECT_SAMPLE_FACTORS Drosophila s5-2 Genotype:w- Luciferase Sample Description=fat body tissues of 4DO larvae collected and homogenized in D PBS without Mg2+, Ca2+; StartVolume=50 µL; Concentration=10 fat bodies/50µL SUBJECT_SAMPLE_FACTORS Drosophila s5-3 Genotype:w- Luciferase Sample Description=fat body tissues of 4DO larvae collected and homogenized in D PBS without Mg2+, Ca2+; StartVolume=50 µL; Concentration=10 fat bodies/50µL SUBJECT_SAMPLE_FACTORS Drosophila s1-1 Genotype:dTorKO Luciferase Sample Description=fat body tissues of 4DO larvae collected and homogenized in D PBS without Mg2+, Ca2+; StartVolume=50 µL; Concentration=10 fat bodies/50µL SUBJECT_SAMPLE_FACTORS Drosophila s1-2 Genotype:dTorKO Luciferase Sample Description=fat body tissues of 4DO larvae collected and homogenized in D PBS without Mg2+, Ca2+; StartVolume=50 µL; Concentration=10 fat bodies/50µL SUBJECT_SAMPLE_FACTORS Drosophila s1-3 Genotype:dTorKO Luciferase Sample Description=fat body tissues of 4DO larvae collected and homogenized in D PBS without Mg2+, Ca2+; StartVolume=50 µL; Concentration=10 fat bodies/50µL SUBJECT_SAMPLE_FACTORS Drosophila s4-1 Genotype:dTorKO Lipin Sample Description=fat body tissues of 4DO larvae collected and homogenized in D PBS without Mg2+, Ca2+; StartVolume=50 µL; Concentration=10 fat bodies/50µL SUBJECT_SAMPLE_FACTORS Drosophila s4-2 Genotype:dTorKO Lipin Sample Description=fat body tissues of 4DO larvae collected and homogenized in D PBS without Mg2+, Ca2+; StartVolume=50 µL; Concentration=10 fat bodies/50µL SUBJECT_SAMPLE_FACTORS Drosophila s4-3 Genotype:dTorKO Lipin Sample Description=fat body tissues of 4DO larvae collected and homogenized in D PBS without Mg2+, Ca2+; StartVolume=50 µL; Concentration=10 fat bodies/50µL SUBJECT_SAMPLE_FACTORS Drosophila s3-1 Genotype:dTorKO Ctdnep1 Sample Description=fat body tissues of 4DO larvae collected and homogenized in D PBS without Mg2+, Ca2+; StartVolume=50 µL; Concentration=10 fat bodies/50µL SUBJECT_SAMPLE_FACTORS Drosophila s3-2 Genotype:dTorKO Ctdnep1 Sample Description=fat body tissues of 4DO larvae collected and homogenized in D PBS without Mg2+, Ca2+; StartVolume=50 µL; Concentration=10 fat bodies/50µL SUBJECT_SAMPLE_FACTORS Drosophila s3-3 Genotype:dTorKO Ctdnep1 Sample Description=fat body tissues of 4DO larvae collected and homogenized in D PBS without Mg2+, Ca2+; StartVolume=50 µL; Concentration=10 fat bodies/50µL SUBJECT_SAMPLE_FACTORS Drosophila s2-1 Genotype:dTorKO Nep1r1 Sample Description=fat body tissues of 4DO larvae collected and homogenized in D PBS without Mg2+, Ca2+; StartVolume=50 µL; Concentration=10 fat bodies/50µL SUBJECT_SAMPLE_FACTORS Drosophila s2-2 Genotype:dTorKO Nep1r1 Sample Description=fat body tissues of 4DO larvae collected and homogenized in D PBS without Mg2+, Ca2+; StartVolume=50 µL; Concentration=10 fat bodies/50µL SUBJECT_SAMPLE_FACTORS Drosophila s2-3 Genotype:dTorKO Nep1r1 Sample Description=fat body tissues of 4DO larvae collected and homogenized in D PBS without Mg2+, Ca2+; StartVolume=50 µL; Concentration=10 fat bodies/50µL SUBJECT_SAMPLE_FACTORS Drosophila s8-1 Genotype:w- Lipin Sample Description=fat body tissues of 4DO larvae collected and homogenized in D PBS without Mg2+, Ca2+; StartVolume=50 µL; Concentration=10 fat bodies/50µL SUBJECT_SAMPLE_FACTORS Drosophila s8-2 Genotype:w- Lipin Sample Description=fat body tissues of 4DO larvae collected and homogenized in D PBS without Mg2+, Ca2+; StartVolume=50 µL; Concentration=10 fat bodies/50µL SUBJECT_SAMPLE_FACTORS Drosophila s8-3 Genotype:w- Lipin Sample Description=fat body tissues of 4DO larvae collected and homogenized in D PBS without Mg2+, Ca2+; StartVolume=50 µL; Concentration=10 fat bodies/50µL SUBJECT_SAMPLE_FACTORS Drosophila s9-1 Genotype:w- Ctdnep1 Sample Description=fat body tissues of 4DO larvae collected and homogenized in D PBS without Mg2+, Ca2+; StartVolume=50 µL; Concentration=10 fat bodies/50µL SUBJECT_SAMPLE_FACTORS Drosophila s9-2 Genotype:w- Ctdnep1 Sample Description=fat body tissues of 4DO larvae collected and homogenized in D PBS without Mg2+, Ca2+; StartVolume=50 µL; Concentration=10 fat bodies/50µL SUBJECT_SAMPLE_FACTORS Drosophila s9-3 Genotype:w- Ctdnep1 Sample Description=fat body tissues of 4DO larvae collected and homogenized in D PBS without Mg2+, Ca2+; StartVolume=50 µL; Concentration=10 fat bodies/50µL #COLLECTION CO:COLLECTION_SUMMARY Fat body tissues of the indicated genotypes were collected and stored at -80°C CO:COLLECTION_SUMMARY until processed for MS analysis. Quantitative MS analysis was performed on ten 4 CO:COLLECTION_SUMMARY day-old Drosophila larval fat bodies homogenized in 50µl D-PBS (Dulbecco’s CO:COLLECTION_SUMMARY Phosphate Buffered Saline without Mg2+ and Ca2+) by Lipotype using previously CO:COLLECTION_SUMMARY described methods (Grillet et al, 2016). CO:SAMPLE_TYPE Insect tissue #TREATMENT TR:TREATMENT_SUMMARY In this study we did not apply any additional treatment(s) to the samples. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Lipids from fat body tissues of indicated genotypes were extracted using SP:SAMPLEPREP_SUMMARY chloroform and methanol (1). Samples were spiked with lipid class-specific SP:SAMPLEPREP_SUMMARY internal standards prior to extraction. After drying and resuspending in MS SP:SAMPLEPREP_SUMMARY acquisition mixture, lipid extracts were subjected to mass spectrometric SP:SAMPLEPREP_SUMMARY analysis. Sample preparations were done by Lipotype. (1) Sampaio, J. L. et al. SP:SAMPLEPREP_SUMMARY Membrane lipidome of an epithelial cell line. Proceedings of the National SP:SAMPLEPREP_SUMMARY Academy of Sciences 108, 1903–1907 (2011). #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY NA CH:CHROMATOGRAPHY_TYPE None (Direct infusion) CH:INSTRUMENT_NAME none CH:COLUMN_NAME none #ANALYSIS AN:ANALYSIS_TYPE MS AN:LABORATORY_NAME Lipotype #MS MS:INSTRUMENT_NAME Thermo Q Exactive Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS Spectra acquisition Mass spectra were acquired on a hybrid quadrupole/Orbitrap MS:MS_COMMENTS mass spectrometer equipped with an automated nano-flow electrospray ion source MS:MS_COMMENTS in both positive and negative ion mode. Data processing and normalization Lipid MS:MS_COMMENTS identification using LipotypeXplorer (2) was performed on unprocessed (*.raw MS:MS_COMMENTS format) mass spectra. For MS-only mode, lipid identification was based on the MS:MS_COMMENTS molecular masses of the intact molecules. MSMS mode included the collision MS:MS_COMMENTS induced fragmentation of lipid molecules and lipid identification was based on MS:MS_COMMENTS both the intact masses and the masses of the fragments. Prior to normalization MS:MS_COMMENTS and further statistical analysis lipid identifications were filtered according MS:MS_COMMENTS to mass accuracy, occupation threshold, noise and background. Lists of MS:MS_COMMENTS identified lipids and their intensities were stored in a database optimized for MS:MS_COMMENTS the particular structure inherent to lipidomic datasets. Intensity of lipid MS:MS_COMMENTS class-specific internal standards was used for lipid quantification. (2) Herzog, MS:MS_COMMENTS R. et al. A novel informatics concept for high-throughput shotgun lipidomics MS:MS_COMMENTS based on the molecular fragmentation query language. Genome Biol. 12, R8 (2011). #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS pmol MS_METABOLITE_DATA_START Samples s5-1 s5-2 s5-3 s1-1 s1-2 s1-3 s4-1 s4-2 s4-3 s3-1 s3-2 s3-3 s2-1 s2-2 s2-3 s8-1 s8-2 s8-3 s9-1 s9-2 s9-3 Factors Genotype:w- Luciferase Genotype:w- Luciferase Genotype:w- Luciferase Genotype:dTorKO Luciferase Genotype:dTorKO Luciferase Genotype:dTorKO Luciferase Genotype:dTorKO Lipin Genotype:dTorKO Lipin Genotype:dTorKO Lipin Genotype:dTorKO Ctdnep1 Genotype:dTorKO Ctdnep1 Genotype:dTorKO Ctdnep1 Genotype:dTorKO Nep1r1 Genotype:dTorKO Nep1r1 Genotype:dTorKO Nep1r1 Genotype:w- Lipin Genotype:w- Lipin Genotype:w- Lipin Genotype:w- Ctdnep1 Genotype:w- Ctdnep1 Genotype:w- Ctdnep1 CE 14:0 1.44965004920959 CE 15:0 2.95913004875183 2.49397993087769 CE 16:0 7.38338994979858 3.28427004814148 CE 16:1 6.8098201751709 CE 18:1 4.53902006149292 CE 21:0 2.84206008911133 3.85169005393982 DAG 30:0 1.51469695568085 2.55234503746033 DAG 30:1 23.4195003509521 23.6122970581055 21.2190113067627 2.28541898727417 4.70481300354004 3.80560803413391 6.23324775695801 5.32875394821167 6.4435887336731 10.2941055297852 10.6690082550049 11.0880393981934 7.67412805557251 7.87699604034424 6.33600997924805 46.0918922424316 41.3443298339844 44.3018798828125 35.6858100891113 41.5352210998535 38.3769607543945 DAG 30:2 5.31798505783081 5.80100297927856 5.81055116653442 0.961960971355438 2.23608994483948 1.57321000099182 2.26425004005432 1.9951000213623 2.4488799571991 3.73186802864075 4.14188480377197 4.51914310455322 2.30482816696167 13.4929714202881 10.7548942565918 12.3825788497925 13.0320262908936 14.4608221054077 10.5942001342773 DAG 31:0 1.87015104293823 2.03522300720215 DAG 31:1 0.811109662055969 2.33627915382385 1.76759302616119 1.71582400798798 7.50643444061279 7.27066707611084 7.22247982025146 DAG 31:2 1.21148300170898 DAG 32:0 4.33145809173584 DAG 32:1 38.0183601379395 42.1727867126465 38.1941452026367 4.60341739654541 8.2989387512207 6.92571020126343 11.8798847198486 10.949330329895 11.6385498046875 16.1334552764893 19.4628639221191 18.2680473327637 16.7364196777344 14.5435400009155 15.2204189300537 83.8974227905273 74.4491424560547 79.9752197265625 62.8192596435547 73.2731094360352 71.6394958496094 DAG 32:2 25.1957302093506 29.44508934021 28.2749404907227 3.93292093276978 7.22048854827881 6.06566333770752 9.21645355224609 8.86759948730469 8.64992141723633 14.2416000366211 19.0525207519531 16.0841293334961 12.3389797210693 11.3897304534912 12.7045593261719 53.7756385803223 49.5854682922363 51.8749008178711 46.1286582946777 55.7267608642578 50.7076721191406 DAG 32:3 0.667096018791199 1.42289710044861 DAG 33:0 1.60101509094238 DAG 33:1 3.09652614593506 3.92194795608521 1.4440850019455 3.62949800491333 2.86139392852783 2.38875818252563 2.6780161857605 9.00361633300781 7.77742004394531 7.64883995056152 7.63744068145752 DAG 33:2 0.670094013214111 1.67446994781494 DAG 34:0 4.40240001678467 1.47462999820709 4.03118991851807 2.30679988861084 2.98143005371094 2.39755988121033 2.04338002204895 4.64147996902466 4.48464012145996 3.19917011260986 2.96876001358032 3.51119995117188 6.51854991912842 5.97892999649048 5.85311985015869 14.2186002731323 14.4390001296997 13.4333000183105 DAG 34:1 32.3828010559082 35.8147201538086 30.9286994934082 5.85171699523926 10.1926002502441 9.41118144989014 14.6150026321411 13.4569540023804 13.9988260269165 18.2008991241455 22.2350006103516 20.9976005554199 16.6942005157471 17.2392997741699 18.5136203765869 77.5760498046875 66.5080795288086 70.6927032470703 71.9538192749023 81.5927658081055 81.0854034423828 DAG 34:2 25.2001399993896 28.1401309967041 25.6036605834961 5.16370820999146 9.56577014923096 7.83535003662109 10.3999099731445 9.4965705871582 11.0418796539307 17.4326305389404 21.2366199493408 18.6616592407227 14.4731397628784 16.0520896911621 16.9887390136719 59.0449981689453 50.601001739502 50.6416397094727 47.2146186828613 58.5637588500977 53.8400268554688 DAG 34:3 7.31207609176636 7.40305042266846 5.8701491355896 1.12393999099731 2.54718995094299 1.9088499546051 2.09477996826172 2.16160011291504 4.21427297592163 6.1445198059082 4.93234300613403 3.25142002105713 2.87951993942261 3.34794998168945 14.5924863815308 13.8705787658691 11.9694118499756 8.42382907867432 9.71718215942383 12.6143636703491 DAG 35:1 1.32358002662659 2.18476009368896 2.58767700195312 DAG 36:0 5.03794002532959 3.90561008453369 3.18698000907898 3.48483991622925 3.55919003486633 19.3505001068115 20.9937000274658 17.7238998413086 DAG 36:1 1.90327000617981 1.07510995864868 2.03818988800049 DAG 36:2 5.70758819580078 5.83161687850952 4.65442276000977 1.49944543838501 3.10011887550354 2.36774706840515 3.3222599029541 3.44725394248962 3.23935389518738 5.94025135040283 5.57655811309814 4.6728310585022 6.75487995147705 6.72563791275024 8.23481273651123 23.5619812011719 19.264440536499 19.6936302185059 17.6989402770996 18.7373104095459 16.7737503051758 DAG 36:3 0.756558001041412 1.57438004016876 PA 28:0 0.735710978507996 0.66922402381897 0.645128011703491 PA 30:1 0.544128000736237 0.961786031723022 0.913465023040771 0.714539766311646 1.05770802497864 1.07480502128601 1.16129100322723 PA 32:1 1.22352540493011 0.939908027648926 0.973361015319824 0.514868021011353 0.702857375144958 0.984683036804199 0.536539018154144 0.584482431411743 0.76693058013916 1.70863199234009 1.93418049812317 1.82764112949371 3.05293226242065 2.64164686203003 2.6771228313446 PA 32:2 1.65661692619324 1.60591244697571 1.64712619781494 0.609103679656982 0.517126023769379 0.677777051925659 0.911045551300049 1.02504169940948 1.07359004020691 0.836987853050232 0.855290412902832 0.921638965606689 3.10168075561523 3.65376996994019 3.13398599624634 3.21630811691284 2.76672005653381 2.79308795928955 PA 34:0 0.225779995322227 0.234299004077911 0.194589003920555 0.432805001735687 0.22316700220108 0.161682993173599 0.469256013631821 0.632282018661499 0.221699997782707 0.352526009082794 0.381141006946564 0.403481006622314 0.357311010360718 1.1166900396347 0.248125001788139 0.935482025146484 0.681973993778229 0.65012401342392 PA 34:1 0.874877035617828 0.711382865905762 0.787324011325836 0.729408979415894 0.597388982772827 0.663604021072388 0.970032036304474 1.19890701770782 1.0957989692688 0.764052033424377 0.956459999084473 0.964569985866547 1.78003096580505 1.88140296936035 1.70067691802979 3.72959995269775 3.32483792304993 2.9896068572998 PA 34:2 3.10503816604614 2.90283012390137 2.83887124061584 0.339566618204117 0.270629405975342 1.36698198318481 1.27675604820251 1.36369645595551 1.93894851207733 2.33894276618958 2.30953598022461 1.96690535545349 2.12590408325195 2.4745135307312 7.3631157875061 8.0563497543335 6.64465665817261 6.85420083999634 5.75697803497314 6.25792407989502 PA 34:3 1.93406999111176 2.06481003761292 1.94641995429993 0.764437973499298 0.823455989360809 0.807444989681244 1.35878205299377 1.38785994052887 1.6739399433136 1.01266002655029 0.927574992179871 1.17637002468109 3.26149010658264 3.48797011375427 3.03454995155334 3.31133008003235 2.75169992446899 2.48391008377075 PA 36:0 0.522484064102173 PA 36:2 2.33261799812317 1.8715660572052 1.69530594348907 0.99481600522995 0.955168008804321 1.08289301395416 1.43880701065063 1.94507098197937 1.332927942276 1.76403903961182 2.31265115737915 2.40617203712463 8.51006984710693 8.72872066497803 7.6967601776123 6.23260831832886 5.05116987228394 5.780930519104 PA 36:3 1.74942803382874 1.52195799350739 1.29705393314362 0.396617501974106 0.260179996490479 0.834809005260468 0.891943991184235 0.73332405090332 1.18133199214935 1.55280005931854 1.30385994911194 1.12321305274963 1.18893909454346 1.09982407093048 3.36141586303711 3.88234210014343 2.93929004669189 3.86409091949463 2.66805601119995 2.81694912910461 PC 26:0 3.78425002098083 3.59714007377625 5.54931020736694 5.49444007873535 PC 28:0 2.48786902427673 2.5891010761261 1.95589995384216 1.93875002861023 2.41343092918396 2.65346193313599 2.76735305786133 2.33077597618103 2.60352993011475 2.36715197563171 2.12231588363647 9.05193519592285 9.42070007324219 8.05020999908447 9.4082202911377 13.8372402191162 12.0850486755371 PC 28:1 4.24255990982056 4.92142963409424 4.33597993850708 6.64834022521973 6.33675003051758 7.81029987335205 6.16909980773926 6.35130023956299 5.78569984436035 5.84844017028809 5.36964988708496 5.9849100112915 27.0266990661621 24.6889991760254 25.9617004394531 32.5113983154297 34.7151031494141 33.8092994689941 PC 28:2 1.13543999195099 0.983803987503052 1.07602000236511 4.70946979522705 PC 30:0 6.18703603744507 6.49086093902588 6.63247013092041 7.17860889434814 10.1243648529053 8.18293380737305 PC 30:2 6.14745998382568 6.34776020050049 6.11071014404297 0.591373980045319 1.37916004657745 8.20792007446289 7.41128015518188 8.15005970001221 7.78007984161377 7.73407983779907 6.50238990783691 6.91159009933472 6.12972021102905 6.46783018112183 27.4041004180908 25.1348724365234 26.9291801452637 30.064920425415 30.1700000762939 25.8081798553467 PC 32:0 4.3974781036377 PC 32:1 158.8955078125 150.28630065918 143.551300048828 7.0130500793457 17.4365119934082 7.42033004760742 61.1618309020996 53.0857009887695 63.504997253418 89.8092498779297 95.1628952026367 87.8785552978516 75.0792999267578 77.7182006835938 74.3779983520508 296.831695556641 271.005279541016 278.536437988281 273.656860351562 284.322479248047 266.843811035156 PC 32:2 163.936767578125 158.011047363281 157.412078857422 7.87959003448486 18.7034206390381 8.8541898727417 71.3567428588867 63.8668785095215 73.772216796875 110.249725341797 113.820495605469 103.289749145508 91.1777877807617 87.8945617675781 88.0785522460938 323.364196777344 296.556304931641 292.622283935547 239.666412353516 261.565490722656 242.707092285156 PC 32:3 2.92970991134644 3.2772798538208 2.65869998931885 0.828881025314331 1.70747005939484 2.24790406227112 3.12650012969971 2.91355991363525 2.71195006370544 2.41501998901367 2.36529016494751 6.31771993637085 6.66460037231445 9.24624061584473 8.92226982116699 5.80709981918335 5.43801021575928 PC 34:0 3.51192998886108 PC 34:1 94.6994018554688 99.5800018310547 91.8368988037109 4.84104204177856 11.3071985244751 5.66284990310669 46.1016311645508 42.9345016479492 48.412914276123 71.9112014770508 75.8469009399414 74.7119979858398 60.5131988525391 61.0704536437988 62.0434837341309 165.824996948242 156.250213623047 156.082366943359 226.093658447266 228.725997924805 225.243286132812 PC 34:2 140.966506958008 130.857803344727 128.741897583008 10.1937799453735 22.0609569549561 12.394437789917 69.2842254638672 61.2876815795898 73.8129730224609 113.862998962402 112.323806762695 108.773895263672 93.8886032104492 96.9384994506836 95.6197052001953 297.507690429688 273.074890136719 267.916381835938 262.966400146484 264.424163818359 260.483489990234 PC 34:3 35.4024085998535 35.3369216918945 34.8288421630859 3.80960988998413 8.59758567810059 4.54718494415283 16.5669994354248 15.8711996078491 16.7179183959961 34.7751312255859 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42.8526992797852 38.1260986328125 37.4785003662109 39.3949012756348 PC 36:4 13.5448799133301 15.2957801818848 12.3709402084351 0.847801029682159 2.69111704826355 6.02622985839844 6.21337985992432 11.8317499160767 11.6788196563721 10.1323795318604 18.6819190979004 PE 26:0 0.907293975353241 0.841816008090973 0.768060028553009 0.611114978790283 0.685801982879639 0.687461972236633 0.852564990520477 1.01192998886108 0.93123197555542 0.719497978687286 0.639034986495972 0.675559997558594 1.93208003044128 1.71158003807068 1.86442995071411 3.51252007484436 4.52938985824585 3.85155010223389 PE 27:0 0.820303022861481 0.774805009365082 1.50943994522095 1.50091481208801 1.44412994384766 PE 28:0 1.42537999153137 1.40041995048523 1.25094997882843 1.03690195083618 0.8072509765625 1.16612005233765 1.5508279800415 1.83961999416351 1.67294800281525 1.25680303573608 1.07899796962738 1.17156004905701 3.83873105049133 3.30796480178833 3.46039485931396 7.31251001358032 7.41336011886597 7.06270980834961 PE 28:1 2.68909883499146 2.56056213378906 2.40760183334351 2.43979001045227 2.38660001754761 2.8452000617981 2.74703598022461 2.70065808296204 2.61711311340332 2.68734979629517 2.50555086135864 2.66713500022888 9.66012954711914 9.41606998443604 9.19869995117188 15.092529296875 15.9130201339722 15.207329750061 PE 28:2 1.4329400062561 0.989310026168823 1.30685997009277 PE 29:0 1.10856997966766 0.8819540143013 1.09776401519775 0.811933994293213 0.697443008422852 0.933326959609985 0.926297008991241 0.871149957180023 0.727745056152344 1.1655969619751 1.06165301799774 1.25379002094269 3.82349514961243 3.83625507354736 3.44598197937012 3.60189390182495 3.84316611289978 3.84421014785767 PE 29:1 1.05411601066589 0.904650986194611 1.0343930721283 0.523051977157593 0.575874984264374 0.551989018917084 0.683835983276367 0.605827987194061 0.808319985866547 1.84127306938171 1.71159911155701 2.06295204162598 2.94515323638916 3.18206405639648 3.02832102775574 PE 30:0 0.48446398973465 0.846990466117859 0.789436995983124 1.58754003047943 1.7622184753418 1.39709997177124 1.09953236579895 1.0113799571991 4.05874013900757 3.94531011581421 4.11377000808716 6.36370706558228 7.14490699768066 5.95255517959595 PE 30:1 55.5930480957031 55.8328514099121 53.2503242492676 1.50072705745697 2.93939518928528 1.88647997379303 22.0075397491455 21.9555130004883 24.3697700500488 29.5985240936279 31.9303035736084 29.0802783966064 25.8159217834473 26.4981365203857 27.7104015350342 112.365913391113 112.569267272949 110.284744262695 104.058685302734 115.241165161133 105.623649597168 PE 30:2 1.2647899389267 1.42080593109131 1.13612997531891 1.70247900485992 1.53334999084473 1.80640995502472 1.62575995922089 1.68406403064728 1.57158994674683 1.65786004066467 1.53061997890472 1.41688001155853 4.50425481796265 4.09539604187012 4.41051244735718 7.90148735046387 8.59059047698975 8.39724922180176 PE 31:0 2.0855610370636 2.03565287590027 1.67188799381256 0.565685749053955 0.9444300532341 1.04930901527405 1.13662910461426 1.06309413909912 0.875756323337555 0.86821973323822 0.826181888580322 6.98598003387451 7.14617013931274 6.94432020187378 5.31888008117676 5.45004987716675 4.6533899307251 PE 31:1 4.01244592666626 4.11554384231567 3.74756789207458 0.320623010396957 0.688039004802704 0.477094799280167 6.37649774551392 6.15353107452393 7.10084199905396 4.729079246521 4.91244173049927 4.38379764556885 4.58654832839966 4.66801929473877 4.8485279083252 19.237943649292 18.6726398468018 19.2254486083984 22.3340644836426 23.7388153076172 21.850658416748 PE 31:2 0.620361983776093 0.665014028549194 0.754499018192291 1.33922898769379 0.911926031112671 1.26659464836121 PE 32:0 1.61601996421814 1.85336005687714 2.72132992744446 PE 32:1 299.128601074219 297.938995361328 279.115112304688 10.5731506347656 21.4689102172852 13.3903799057007 128.481307983398 127.622901916504 139.820358276367 191.74528503418 203.715805053711 187.323699951172 155.541000366211 159.129257202148 162.28955078125 605.351989746094 585.993774414062 585.53466796875 599.279968261719 661.056518554688 612.232055664062 PE 32:2 100.800598144531 100.951675415039 98.7552947998047 5.34560680389404 12.206919670105 7.35623407363892 53.4045867919922 51.3973121643066 57.0236701965332 78.3899002075195 81.5531997680664 75.5815048217773 62.4648132324219 61.376407623291 64.9177017211914 198.779602050781 192.500854492188 189.806030273438 200.210556030273 225.877395629883 206.894485473633 PE 33:0 5.71435022354126 5.89600038528442 5.87097978591919 5.17449998855591 5.27490711212158 5.43269014358521 PE 33:1 11.7626800537109 12.0874099731445 11.0921497344971 0.712917983531952 1.84560608863831 1.14615499973297 14.5242700576782 13.7234601974487 14.9683294296265 12.1077499389648 12.7366104125977 12.1982402801514 11.5138301849365 11.3256702423096 12.1307697296143 43.254150390625 42.0065231323242 40.6766510009766 48.4744987487793 55.4771003723145 48.4166984558105 PE 33:2 4.35630035400391 5.02040004730225 4.3728199005127 0.627110004425049 1.12918198108673 1.11329400539398 7.35023975372314 7.19945001602173 7.7859001159668 5.26880979537964 5.693039894104 5.22680997848511 4.23273372650146 4.61917972564697 4.35509014129639 15.8708610534668 15.483829498291 15.1141004562378 18.3995800018311 20.279369354248 18.6153202056885 PE 34:1 300.535003662109 295.309997558594 259.457000732422 22.1424999237061 49.8172988891602 30.2401008605957 150.573852539062 142.630004882812 161.875045776367 234.817993164062 228.64599609375 218.039993286133 190.205993652344 182.050994873047 184.572006225586 686.47998046875 652.994995117188 655.265991210938 642.265014648438 738.830017089844 657.051025390625 PE 34:2 335.131988525391 335.880004882812 321.414978027344 20.5607395172119 46.3118019104004 27.8797492980957 159.674545288086 154.126235961914 172.723098754883 262.213104248047 276.013885498047 262.1875 212.786407470703 217.266464233398 219.83869934082 692.031982421875 671.087036132812 663.06298828125 633.554016113281 696.127014160156 651.349243164062 PE 34:3 50.3358345031738 49.1844482421875 46.6760597229004 4.41388082504272 10.1208477020264 5.90896606445312 22.661470413208 21.1862106323242 23.9737396240234 46.8885803222656 47.7165985107422 45.8275718688965 32.802490234375 31.7123012542725 32.6594276428223 87.8567810058594 82.6734008789062 82.9763259887695 91.3730850219727 100.600547790527 94.8920669555664 PE 34:4 1.40500998497009 1.5769499540329 1.2858099937439 0.560348987579346 0.566538989543915 0.598780989646912 1.15464997291565 1.36423003673553 1.3666900396347 0.786387026309967 0.90227997303009 0.822393000125885 2.28467988967896 1.87957000732422 1.88445997238159 2.95019006729126 3.2055299282074 2.77869009971619 PE 35:1 10.8378610610962 10.5395450592041 10.0470056533813 1.32395303249359 2.73790502548218 1.56951904296875 8.74447822570801 8.3439302444458 9.46035957336426 10.2656116485596 10.2568473815918 10.1013889312744 9.56612682342529 9.67370510101318 10.1861190795898 31.5023975372314 30.8446083068848 30.4647235870361 28.7150440216064 33.0757942199707 29.8387031555176 PE 35:2 7.45920610427856 7.44047260284424 7.09767007827759 1.00234603881836 1.98493909835815 1.34684801101685 8.90042209625244 8.71077156066895 9.65910720825195 8.32108688354492 8.32175731658936 8.44474124908447 7.2576756477356 7.14926099777222 7.20296192169189 20.8918190002441 19.5571002960205 19.220251083374 21.9960994720459 24.572790145874 22.3415298461914 PE 35:3 1.16925001144409 1.41081500053406 1.20408701896667 0.267978996038437 1.52543997764587 1.51463997364044 1.44693005084991 1.6385999917984 1.54837000370026 1.33651995658875 1.21586000919342 1.15562999248505 1.23341906070709 3.05975008010864 2.68401002883911 2.69713020324707 3.06402993202209 3.66771006584167 3.73643398284912 PE 36:1 12.3908205032349 12.940149307251 11.6898002624512 1.97527503967285 4.33547306060791 2.79062509536743 7.80023002624512 6.68696022033691 7.87040996551514 13.4063701629639 12.7614498138428 12.4557600021362 10.4805698394775 10.0673799514771 10.0234498977661 30.4330101013184 29.7210311889648 29.847900390625 32.855598449707 37.2516021728516 32.8083992004395 PE 36:2 175.007736206055 173.883911132812 165.442749023438 12.3807792663574 28.1287307739258 16.9843502044678 93.2474670410156 87.671501159668 100.547180175781 160.841827392578 162.541427612305 155.464111328125 136.59912109375 135.587539672852 137.964981079102 439.144927978516 433.703399658203 432.226165771484 327.08251953125 370.390014648438 337.303619384766 PE 36:3 69.8315582275391 67.3290710449219 63.3853988647461 6.85388994216919 15.9044399261475 9.87448024749756 29.3545246124268 27.485200881958 30.7138004302979 66.4160995483398 69.4587020874023 64.907600402832 49.532154083252 46.8965301513672 49.2636985778809 130.659362792969 125.166976928711 123.367668151855 110.163009643555 121.598999023438 114.463996887207 PE 36:4 10.8456802368164 9.92951011657715 9.65153026580811 1.5744891166687 3.98729300498962 2.22722291946411 3.80087995529175 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90.8233947753906 87.1208038330078 4.75914621353149 9.24442100524902 5.76642513275146 26.8935203552246 24.5427074432373 27.8533344268799 44.639404296875 48.1497955322266 45.3599739074707 33.0334625244141 34.4951286315918 33.1373558044434 213.825698852539 197.103378295898 193.813995361328 128.018844604492 134.69596862793 134.917526245117 PI 34:3 19.1869297027588 17.6991500854492 16.7599086761475 1.80126190185547 3.07614707946777 1.9217209815979 5.93745708465576 5.76741981506348 6.01855993270874 11.8028898239136 13.3341093063354 11.7893390655518 8.50406932830811 8.63901996612549 7.92311096191406 32.5153121948242 29.6537399291992 28.6803703308105 26.9097099304199 29.2321891784668 28.9721984863281 PI 34:4 2.29921889305115 1.75660002231598 2.02531504631042 0.677330017089844 0.856517970561981 0.683463990688324 1.29454207420349 1.56462800502777 1.4794100522995 1.11089706420898 0.957327961921692 0.939052999019623 2.90443992614746 2.63585305213928 2.61809992790222 2.63930010795593 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DAG GL MEM 33 0 0 DAG 33:1 DAG GL MEM 33 1 0 DAG 33:2 DAG GL MEM 33 2 0 DAG 34:0 DAG GL MEM 34 0 0 DAG 34:1 DAG GL MEM 34 1 0 DAG 34:2 DAG GL MEM 34 2 0 DAG 34:3 DAG GL MEM 34 3 0 DAG 35:1 DAG GL MEM 35 1 0 DAG 36:0 DAG GL MEM 36 0 0 DAG 36:1 DAG GL MEM 36 1 0 DAG 36:2 DAG GL MEM 36 2 0 DAG 36:3 DAG GL MEM 36 3 0 PA 28:0 PA GPL MEM 28 0 0 PA 30:1 PA GPL MEM 30 1 0 PA 32:1 PA GPL MEM 32 1 0 PA 32:2 PA GPL MEM 32 2 0 PA 34:0 PA GPL MEM 34 0 0 PA 34:1 PA GPL MEM 34 1 0 PA 34:2 PA GPL MEM 34 2 0 PA 34:3 PA GPL MEM 34 3 0 PA 36:0 PA GPL MEM 36 0 0 PA 36:2 PA GPL MEM 36 2 0 PA 36:3 PA GPL MEM 36 3 0 PC 26:0 PC GPL MEM 26 0 0 PC 28:0 PC GPL MEM 28 0 0 PC 28:1 PC GPL MEM 28 1 0 PC 28:2 PC GPL MEM 28 2 0 PC 30:0 PC GPL MEM 30 0 0 PC 30:2 PC GPL MEM 30 2 0 PC 32:0 PC GPL MEM 32 0 0 PC 32:1 PC GPL MEM 32 1 0 PC 32:2 PC GPL MEM 32 2 0 PC 32:3 PC GPL MEM 32 3 0 PC 34:0 PC GPL MEM 34 0 0 PC 34:1 PC GPL MEM 34 1 0 PC 34:2 PC GPL MEM 34 2 0 PC 34:3 PC GPL MEM 34 3 0 PC 34:4 PC GPL MEM 34 4 0 PC 36:2 PC GPL MEM 36 2 0 PC 36:3 PC GPL MEM 36 3 0 PC 36:4 PC GPL MEM 36 4 0 PE 26:0 PE GPL MEM 26 0 0 PE 27:0 PE GPL MEM 27 0 0 PE 28:0 PE GPL MEM 28 0 0 PE 28:1 PE GPL MEM 28 1 0 PE 28:2 PE GPL MEM 28 2 0 PE 29:0 PE GPL MEM 29 0 0 PE 29:1 PE GPL MEM 29 1 0 PE 30:0 PE GPL MEM 30 0 0 PE 30:1 PE GPL MEM 30 1 0 PE 30:2 PE GPL MEM 30 2 0 PE 31:0 PE GPL MEM 31 0 0 PE 31:1 PE GPL MEM 31 1 0 PE 31:2 PE GPL MEM 31 2 0 PE 32:0 PE GPL MEM 32 0 0 PE 32:1 PE GPL MEM 32 1 0 PE 32:2 PE GPL MEM 32 2 0 PE 33:0 PE GPL MEM 33 0 0 PE 33:1 PE GPL MEM 33 1 0 PE 33:2 PE GPL MEM 33 2 0 PE 34:1 PE GPL MEM 34 1 0 PE 34:2 PE GPL MEM 34 2 0 PE 34:3 PE GPL MEM 34 3 0 PE 34:4 PE GPL MEM 34 4 0 PE 35:1 PE GPL MEM 35 1 0 PE 35:2 PE GPL MEM 35 2 0 PE 35:3 PE GPL MEM 35 3 0 PE 36:1 PE GPL MEM 36 1 0 PE 36:2 PE GPL MEM 36 2 0 PE 36:3 PE GPL MEM 36 3 0 PE 36:4 PE GPL MEM 36 4 0 PE 37:1 PE GPL MEM 37 1 0 PE 37:2 PE GPL MEM 37 2 0 PE 38:1 PE GPL MEM 38 1 0 PE 38:2 PE GPL MEM 38 2 0 PE 38:3 PE GPL MEM 38 3 0 PG 30:0 PG GPL MEM 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