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Kirschbaum"],"technology_type":["Mass Spectrometry","Data-independent acquisition","Bottom-up proteomics"],"software":[""],"submitter_keywords":[""],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD072869"],"tissue":["Hepatocyte","Cell Culture"],"sample_protocol":["For knockdown of StARD phospholipid transfer proteins in HepG2 cells, we used ON-TARGETplus SMART pool siRNA specifically targeting STARD2, STARD7 and STARD10 (Dharmacon). The siRNA pools contained the following target sequences: STARD2: GGAAGUGGCAUUUCGUUCA, UGAUCCGGGUGAAGCAAUA, CGAUCGAGAGUGACGGCAA, CAAACUGGAUGACGUGCCA; STARD7: CAUCGAUUGUGCAGCGCUU, CCUCUGAGCGAAAGAACGA, UUAAUGAGAUGAAGCGGUU, CCAAUGUACUCACGGGAUU; STARD10: GGAAAGACUUGGUCCGAGC, GACAUUGAGUACCGCAAGA, AGAGCAGAGAGGAGCGGAU, GCCCGAUGACCAAGACUUU. HepG2 cells were seeded into 12-well plates at a seeding density of 10^5 cells per well. After overnight incubation in complete medium (DMEM/F-12 medium supplemented with non-essential amino acids, 10 % FBS), the medium was replaced by 1 mL of transfection solution containing siRNA at a final concentration of 50 nM and 4 µL DharmaFECT 4 transfection reagent per well. According to the manufacturer’s instructions, siRNA and transfection reagent were first incubated in a 200 µL volume for 20 min in serum-free medium before 800 µL complete medium was added. The transfections were performed in six replicates for each protein and controls. Control cells were incubated with transfection reagent under identical conditions without adding siRNA. Cells were incubated with transfection solution at 37 °C with 5 % CO2 for 72 h. Cells were washed twice with PBS, harvested by cell scraping, and prepared for lipid analysis and proteomics.  For proteomics, ca. 6 x 10^5 cells were lyzed in 46 µL lysis buffer (5 % SDS in 50 mM Tris-HCl, pH 7.4) through sonication. After clarification by centrifugation (13,000 × g, 8 min), the supernatant was transferred into a new tube. 2 µL of TCEP (120 mM in water) was added and the reduction was allowed to proceed for 15 min at 55 °C. After incubation with 2 µL iodoacetamide (250 mM in 50 mM Tris-HCl) for 10 min at room temperature, the solution was acidified with 10 µL phosphoric acid (12 % in water). 350 µL wash buffer (100 mM Tris-HCl in 90 % methanol) was added, and the solution was applied to a miniprep column. Proteins were bound to the column by centrifugation (4000 × g, 30 s). The column was washed four times with 400 µL wash buffer through centrifugation and dried by a final centrifugation at 4000 × g for 1 min before being transferred into a new tube. 125 µL digestion buffer was applied to the column (50 mM Tris containing a total of 2 µg trypsin per column). Proteins were digested overnight at 37 °C. Peptides were eluted sequentially with 50 mM Tris, 0.2 % formic acid in water, and 50 % acetonitrile in water through centrifugation (4000 × g, 1 min; 80 µL per elution). The eluates were combined and dried in a centrifugal evaporator. The dried peptides were resuspended in 0.1 % formic acid and diluted to a final concentration of 100 ng/µL. Purified peptides were separated via nanoflow reversed-phase liquid chromatography (nanoElute 2, Bruker Daltonics) with a 30 min gradient. Peptides (50 ng, 0.5 µL injection volume) were loaded directly onto a 25 cm × 75 µm column packed with 1.7 µm C18 beads (pore size 120 Å; Aurora Ultimate CSI, IonOpticks) and separated at a flow rate of 250 nL/min and a column temperature of 50°C. Mobile phase A consisted of 0.1% formic acid in water, and mobile phase B consisted of 0.1% formic acid in acetonitrile (v/v; Fisher Scientific, LC-MS grade). The gradient was as follows: 2–23% B over 18 min, 23–35% B over 4 min, and 35–90% B over 4 min, followed by a 4-min wash at 90% B. Eluting peptides were infused into a TIMS quadrupole time-of-flight mass spectrometer (timsTOF Ultra, Bruker Daltonics) equipped with an electrospray ion source (CaptiveSpray, Bruker Daltonics). Source parameters were set to a capillary voltage of 4,500 V, a dry gas flow of 3.0 L min⁻¹, and a dry gas temperature of 180 °C. Data was acquired in dia-PASEF mode over an m/z range of 400–1,000 and an ion mobility range of 1/K₀ = 0.64–1.45 Vs/cm2 (Table S4). The TIMS analyzer was operated at a 100% duty cycle with 100 ms accumulation and 100 ms ramp times (acquisition cycle time = 0.96 s). Collision energies were linearly stepped from 20 to 59 eV as a function of increasing ion mobility."],"repository":["Pride"],"quantification_method":[""],"modification":[""],"data_protocol":["dia-PASEF raw files were processed using the “DIA_SpecLib_Quant_diaPASEF” workflow in FragPipe v.23.1. Pseudo-MS/MS spectra were generated using diaTRACER v.1.3.3 and searched using MSFragger v.4.3. Searches were performed against the Homo sapiens UniProt reference proteome (UP000005640; reviewed entries only) supplemented with common contaminants and decoys (downloaded November 2025). Precursor and fragment mass tolerances were set to 20 ppm, and the isotope error window was set to 0/1/2. Mass calibration and parameter optimization were enabled. Enzymatic specificity was set to “stricttrypsin”, allowing up to two missed cleavages. Carbamidomethylation of cysteine was specified as a fixed modification, while methionine oxidation, protein N-terminal acetylation, and pyro-Glu/Gln formation at peptide N-termini were included as variable modifications (maximum of three variable modifications per peptide). MSBooster and Percolator were used to predict retention time and MS/MS spectra, and to rescore peptide–spectrum matches (PSMs). PSMs were filtered to a 1% false-discovery rate (FDR) and combined with the pseudo-MS/MS spectra using EasyPQP v.0.1.52 to generate a spectral library for DIA quantification (139,376 precursors mapping to 8,002 protein groups). dia-PASEF runs were quantified using DIA-NN v.1.8.2 beta 8 with the spectral library generated in FragPipe. Mass accuracy settings for MS1 and MS2, as well as the scan window, were determined automatically by DIA-NN based on the first run in the experiment. Precursor and protein group identifications were filtered at 1% FDR. Protein inference was disabled, retaining protein group assignments from the input spectral library. Quantification was performed using fixed-width peak-centre integration. Cross-run normalisation was performed using DIA-NN's default RT-dependent method, and protein group quantities were calculated using the MaxLFQ algorithm. Quantification matrices for precursors, protein groups, and gene groups were exported with 1% FDR filtering. The abundance of STARD2, STARD7 and STARD10 were quantified in six replicates per condition (control and three siRNA knockdown conditions)."],"omics_type":["Proteomics"],"labhead":["Carol Robinson"],"instrument_platform":[""],"labhead_affiliation":["Kavli Institute for Nanoscience Discovery, University of Oxford"],"submission_type":["PARTIAL"],"species":["Homo Sapiens (human)"],"publication":["Not available"],"submitter_mail":["carla.kirschbaum@charite.de"],"submitter_affiliation":["Charité - Universitätsmedizin Berlin"],"submitter_country":["Germany"],"additional_accession":[]},"is_claimable":false,"name":"siRNA-mediated knockdown of StARD phospholipid transfer proteins in HepG2 cells","description":"We used siRNA to individually knock down the human phospholipid transfer proteins STARD2, STARD7 and STARD10 in HepG2 cells and measured the impact on the cellular phospholipid metabolism. We used DIA proteomics to quantify the selective reduction in protein expression levels.","dates":{"publication":"2026-07-21","submission":"2026-01-10"},"accession":"PXD072869","cross_references":{"TAXONOMY":["NEWT:330879","NEWT:377960","NEWT:1131","NEWT:35795","NEWT:2042546","NEWT:259447","NEWT:295546","NCBITaxon:2719036","NEWT:1334565","NEWT:625351","NEWT:258594","NCBITaxon:1280","NEWT:112503","NEWT:1129","NEWT:309807","NEWT:59753","NEWT:89184","NEWT:309800","NEWT:281395","NEWT:247480","NEWT:1211601","NEWT:876138","NEWT:44271","NEWT:193516","NEWT:2315688","NEWT:592321","NEWT:1073882","NEWT:6819","NEWT:112509","NEWT:475174","NEWT:111205","NEWT:1599","NEWT:428386","NEWT:1117","NEWT:498257","NEWT:10036","NEWT:32264","NEWT:12215","NEWT:1590","NEWT:10034","NEWT:33114","NEWT:1596","NEWT:661410","NEWT:638632","NEWT:224326","NEWT:376619","NEWT:1748418","NCBITaxon:79857","NEWT:1096976","NEWT:3765","NEWT:3767","NEWT:1589","NEWT:135622","NEWT:4615","NEWT:672127","NEWT:35786","NEWT:1580","NEWT:399784","NEWT:96731","NEWT:656064","NEWT:1582","NEWT:3760","NEWT:1584","NEWT:36630","NEWT:1585","NEWT:383379","NEWT:381198","NEWT:418106","NCBITaxon:12721","NEWT:10029","NEWT:913645","NEWT:2492962","NEWT:641809","NEWT:1392488","NEWT:38815","NEWT:317447","NEWT:4688","NEWT:111225","NEWT:7719","NEWT:5530","NEWT:868565","NEWT:135674","NEWT:1562698","NEWT:79329","NEWT:30069","NEWT:12637","NEWT:59729","NEWT:2164133","NEWT:295105","NEWT:108061","NEWT:60711","NEWT:224308","NEWT:3347","NEWT:49928","NEWT:3349","NEWT:160621","NEWT:2502409","NEWT:212790","NEWT:110368","NEWT:55350","NEWT:1916073","NEWT:1310161","NEWT:77133","NEWT:145481","NEWT:1310165","NEWT:29058","NCBITaxon:79824","NEWT:1912919","NEWT:410658","NEWT:5516","NEWT:44688","NEWT:44689","NEWT:370038","NEWT:498211","NEWT:345076","NEWT:347256","NEWT:5518","NEWT:398007","NEWT:1527468","NEWT:498217","NEWT:498216","NEWT:11320","NEWT:246196","NEWT:246197","NEWT:145458","NEWT:1310173","NEWT:200795","NEWT:37919","NEWT:2579791","NEWT:44685","NEWT:161934","NEWT:156587","NEWT:1148","NEWT:5508","NEWT:3329","NEWT:419481","NEWT:5507","NEWT:260499","NEWT:410661","NEWT:329854","NEWT:1140","NCBITaxon:2157","NEWT:2066563","NEWT:2649329","NEWT:1143","NEWT:1287689","NEWT:1094343","NEWT:1462472","NEWT:1336795","NEWT:644042","NEWT:150808","NEWT:1182590","NEWT:3712","NEWT:3711","NEWT:3714","NCBITaxon:37162","NEWT:270643","NEWT:3715","NEWT:1538","NEWT:2065263","NEWT:177437","NEWT:717966","NEWT:10891","NEWT:10418","NEWT:118698","NEWT:1616117","NEWT:118696","NEWT:34865","NEWT:52283","NEWT:28174","NEWT:284812","NEWT:8175","NEWT:43330","NEWT:1603293","NEWT:44664","NEWT:43346","NEWT:3702","NEWT:1245466","NEWT:3704","NEWT:244366","NEWT:1246791","NEWT:176597","NEWT:118694","NEWT:2850","NEWT:118691","NEWT:34871","NEWT:9006","NEWT:33548","NEWT:52299","NEWT:8165","NEWT:96794","NEWT:3708","NEWT:332648","NEWT:44670","NEWT:926571","NEWT:536231","NEWT:376219","NEWT:219813","NEWT:1510","NEWT:460519","NEWT:1515","NEWT:572307","NEWT:1432138","NEWT:1424507","NEWT:1194599","NEWT:272844","NEWT:1348799","NEWT:33995","NEWT:55784","NEWT:1303443","NEWT:9483","NEWT:485","NEWT:56636","NEWT:2709072","NEWT:2499525","NEWT:2853422","NEWT:1122984","NEWT:487","NEWT:1538553","NEWT:1679718","NEWT:480","NEWT:67767","NEWT:45972","NEWT:46835","NEWT:109757","NEWT:582580","NEWT:1185650","NEWT:216778","NEWT:294607","NEWT:1502","NEWT:128017","NEWT:2836","NEWT:376686","NEWT:237610","NEWT:95486","EFO:0001352","NEWT:119562","NEWT:1883446","NEWT:1233435","NEWT:118223","NEWT:11757","NEWT:987059","NEWT:109760","NEWT:29031","NEWT:317047","NEWT:1509","NEWT:491","NEWT:460523","NEWT:45954","NEWT:235443","NEWT:1579","NEWT:1757312","NEWT:85694","NEWT:109786","NEWT:1922344","NEWT:108458","NEWT:5936","NEWT:85692","NEWT:321967","NEWT:320637","NEWT:1078283","NEWT:3750","NEWT:983964","NEWT:11706","NEWT:32644","NEWT:527796","NEWT:279010","NEWT:499175","NEWT:109779","NEWT:3745","NEWT:1715989","NCBITaxon:4751","NEWT:3747","NEWT:1116234","NEWT:415978","NEWT:1255228","NEWT:410289","NEWT:191701","NEWT:373153","NEWT:1640278","NEWT:9054","NEWT:472","NEWT:940825","NEWT:29459","NEWT:529507","NEWT:1071661","NEWT:28128","NEWT:470","NEWT:41122","NEWT:5911","NEWT:1328300","NEWT:748693","NCBITaxon:50557","NEWT:39251","NEWT:29491","NEWT:101841","NEWT:446","NEWT:153481","NEWT:2014887","NEWT:33952","NEWT:11723","NEWT:445","NEWT:153009","NEWT:261756","NEWT:166314","NEWT:166313","NEWT:63366","NEWT:63367","NEWT:29488","NEWT:215402","NEWT:1547","NEWT:1290454","NEWT:3726","NEWT:906914","NEWT:81231006","NEWT:30455","NEWT:28141","NEWT:9031","NEWT:27292","NEWT:1050722","NEWT:3728","NEWT:108931","NEWT:1293497","NEWT:1055524","NEWT:548","NEWT:150475","NEWT:267872","NEWT:381666","NEWT:172269","NEWT:9534","NEWT:2126348","NEWT:5180","NEWT:256737","NEWT:9541","NEWT:8694","NEWT:33936","NEWT:8692","NEWT:542","NEWT:2903","NEWT:185579","NEWT:872227","NEWT:13076","NEWT:2587410","NEWT:207340","NEWT:1006581","NEWT:8673","NEWT:33940","NEWT:2126337","NEWT:550","NEWT:10804","NEWT:33941","NEWT:554","NEWT:451516","NEWT:552","NEWT:1325291","NEWT:42410","NEWT:36185","NEWT:1054211","N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