{"database":"Pride","file_versions":[{"headers":{"Content-Type":["application/json"]},"body":{"files":{"Txt":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD074138/checksum.txt"],"Raw":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD074138/N2_2.raw.quant.md5","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD074138/L1.raw.quant","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD074138/R2_2.raw.quant.md5","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD074138/R2_2.raw.md5","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD074138/N2_2.raw.md5","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD074138/L3.raw.quant.md5","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD074138/L1.raw.quant.md5","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD074138/N1_2.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD074138/N1_2.raw.md5","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD074138/L1.raw.md5","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD074138/L1.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD074138/R2_2.raw.quant","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD074138/L2.raw.md5","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD074138/L3.raw.md5","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD074138/L2.raw.quant","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD074138/L3.raw.quant","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD074138/N1_2.raw.quant","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD074138/N1_2.raw.quant.md5","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD074138/N2_2.raw.quant","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD074138/N2_2.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD074138/L3.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD074138/L2.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD074138/L2.raw.quant.md5","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD074138/R2_2.raw"],"Other":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD074138/search.zip"]},"type":"primary"},"statusCodeValue":200,"statusCode":"OK"}],"scores":null,"additional":{"labhead_mail":["altas_zhang@163.com"],"submitter":["yonghui chen"],"technology_type":["Mass Spectrometry","Data-independent acquisition"],"software":["Not available"],"submitter_keywords":["Mouse bone marrow‑derived macrophages (bmdms)，5‑ht2b receptor，bw723c86，transwell assay，lc-msms"],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD074138"],"tissue":["Bone Marrow"],"sample_protocol":["1. Protein extraction(1) A proper number of samples were weighed and transferred into 2 mL centrifuge tubes. Steel balls, lysis solution containing 8M Urea/50mMTris-HCL and Roche cocktail with 1X final concentration were added into the centrifuge tubes. The tubes were placed on ice for 5 min. (2) Samples were homogenated using a tissue lyser (60Hz, 2 min) followed by 15 min centrifugation (20,000×g, 4°C). The supernatant was collected. (3) DTT was added to obtain a final concentration of 10 mM, and the tubes were placed in 37℃ water bath for 1 hour. (4) IAA was added to obtain a final concentration of 20 mM, and the tubes were placed away from light for 30 min. 2. Quality control of protein extraction(1) The Bradford method for protein quantitation 0, 2, 4, 6, 8, 12, 14, 16, 18μL standard protein solution (0.2μg/μLBSA) was added into the 96-well plate successively, and then 20, 18, 16, 14, 12, 10, 8, 6, 4, 2 μL pure water was added successively. After mixing thoroughly, 180μL Coomas bright Blue G250 quantitative working solution was added to each well. A microplate reader was used to measure the absorption value at 595nm (OD595), and the linear standard curve was created according to OD595 and protein concentration. 180 μL quantitative working solution was added to 20 μL protein solution of the sample, and the OD595 of the sample was read. The protein concentration of the sample was calculated according to the standard curve and OD595 of the sample. (2) SDSPAGE 10 μg protein was taken from each sample, and the proper volume of loading buffer was added. The mixture was heated at 95℃ for 5 min and centrifuged at 20,000g for 5min. The supernatant was added to the sample well of 4-12% SDS polyacrylamide gel and then subjected to 80V constant pressure electrophoresis for 20 min, followed by 120V constant pressure electrophoresis for 60 min. After electrophoresis, the gel was stained, decolorized and taken out to take photos. 3. Protein digestion(1) 150 μg protein was taken from each sample. (2) 3 μg trypsin was added to obtain a ratio of protein: enzyme =50:1, and the samples were incubated for 14-16h at 37℃. (3) The enzymatically digested peptides were desalted using Waters solid phase extraction cartridges and then vacuum dried. (4) The dried peptide fractions were redissolved in pure water and stored at -20°C.4. Reversed-Phase (HpH) High-Pressure Liquid Chromatography (HPLC) Fractionation Equal amounts of peptide from each sample were mixed, and then diluted with solvent A (5% ACN, pH 9.8) and injected into the column. The peptide mixture was fractionated using a 3.5 μm 4.6x150 mm Agilent ZORBAX 300Extend- C18 column on Thermo Scientific UltiMate™ 3000 Binary Rapid Separation System. The gradient elution was performed at a flow rate of 0.3 mL/min: 5 to 21% solvent B (97% ACN, pH 9.8) in 38 min, 21.5% to 40% solvent B in 20 min, 40% to 90% solvent B in 2 min, 90% solvent B for 3 min, and 5% solvent B equilibrated for 10 min. The elution peaks were monitored at 214 nm and fractions were collected every minute. The fractions were combined according to chromatograms of the elution peaks. Ten fractions were obtained  and then freeze-dried.5. Quantitative detection by Nano-LC-MS/MSMoble phases A (100% water, 0.1% formic acid) and B (80% acetonitrile) were prepared, and the dried peptide samples were reconstituted with 0.1% formic acid and centrifuged at 20,000 g for 10 min. Separation was performed by a Thermo Scientific Vanquish Neo UHPLC liquid phase system. The samples were fed onto a ES906 HPLC column from 150mm with an 8 min gradient at a flow rate of 2.5 μl/min, and were separated withthe following effective gradient: 0-4 min, 4% mobile phase B linearly ramping up to 25%; 4-6.9 min, mobile phase B linearly ramping up from 25% to 35%; 6.9-7.3 min, mobile phase B linearly ramping up from 35% to 99%; From 7.3-8.0min, mobile phase B was maintained at 99%.The liquidphase separated peptides were transferred to a Thermo Astral mass spectrometer for acquisition in DIA(Data Independent Acquisition) mode. The main parameters were set as follows: normalized collision energy of 25%, default charge state of 2, resolution of 240,000, scanning every 06 s, scanning range of 380-980 m/z, and mass spectrometry AGC setting of 500%. Fragmentation ion scans were recorded with a maximum scan time of 3 ms, and we used 300 2-Th scanning windows from 380 to 980 m/z."],"repository":["Pride"],"quantification_method":["Not available"],"modification":[""],"data_protocol":["DIANN was used to analyze the DIA data with libraryfree method. MS/MS data were searched against protein sequences which were downloaded from Uniprot database with the following settings: enzyme: Trypsin/P; maximum missed cleavages: 2; fixed modification: carbamidomethyl (C); variable modifications: oxidation (M) and acetyl (protein N-term); precursor mass tolerance: 20 ppm; fragment mass tolerance: 0.05Da. The results were filtered by 1% FDR, and only those protein groups which passed this filter criteria were used in downstream analysis"],"omics_type":["Proteomics"],"labhead":["yueman Zhang"],"instrument_platform":[""],"submission_type":["PARTIAL"],"labhead_affiliation":["Department of Anesthesiology, Clinical Research Center, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China"],"species":["Mus Musculus (mouse)"],"publication":["Not available"],"submitter_mail":["cyhgordon@163.com"],"submitter_affiliation":["Renji Hospital, Shanghai Jiao Tong University School of Medicine‌"],"submitter_country":["China"],"additional_accession":[]},"is_claimable":false,"name":"Proteomic Profiling of the 5‑HT2B Receptor Agonist BW723C86 on Migration and Signaling in Mouse Bone Marrow‑Derived Macrophages","description":"Mouse Bone marrow-derived macrophages (BMDMs) and HT22 neuronal cells (subjected to 2-hour oxygen-glucose deprivation, OGD-2h) were used in this study. To investigate cell migration, a Transwell assay was performed. For pharmacological intervention, cells were treated with either the serotonin receptor 2B (5-HT2B) agonist BW723C86 or dimethyl sulfoxide (DMSO) as a vehicle control. Following treatment, global protein expression profiling was performed using mass spectrometry-based quantitative proteomics.","dates":{"publication":"2026-07-04","submission":"2026-02-06"},"accession":"PXD074138","cross_references":{"TAXONOMY":["NEWT:330879","NEWT:108018","NEWT:377960","NEWT:1131","NEWT:35795","NEWT:2042546","NEWT:259447","NEWT:295546","NCBITaxon:2719036","NEWT:1334565","NEWT:625351","NEWT:258594","NCBITaxon:1280","NEWT:1126","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:1367493","NEWT:592321","NEWT:1073882","NEWT:6819","NEWT:46429","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:2736291","NEWT:661410","NEWT:638632","NEWT:224326","NEWT:294671","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:656061","NEWT:37965","NEWT:656064","NEWT:1582","NEWT:3760","NEWT:1584","NEWT:2086595","NEWT:36630","NEWT:1585","NEWT:383379","NEWT:381198","NEWT:418106","NCBITaxon:12721","NEWT:166340","NEWT:10029","NEWT:913645","NEWT:2492962","NEWT:641809","NEWT:44250","NEWT:1392488","NEWT:38815","NEWT:317447","NEWT:4688","NEWT:3357","NEWT:111225","NEWT:7719","NEWT:29073","NEWT:5530","NEWT:868565","NEWT:135674","NEWT:1562698","NEWT:79329","NEWT:30069","NEWT:12637","NEWT:59729","NEWT:2164133","NEWT:295105","NEWT:2767846","NEWT:108061","NEWT:60711","NEWT:2020713","NEWT:224308","NEWT:3347","NEWT:49928","NEWT:3349","NEWT:160621","NEWT:2502409","NEWT:212790","NEWT:110368","NEWT:55350","NEWT:1916073","NEWT:1310161","NEWT:1230383","NEWT:77133","NEWT:145481","NEWT:1310165","NEWT:29058","NCBITaxon:79824","NEWT:468822","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:342808","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:99802","NEWT:1287689","NEWT:1094343","NEWT:1462472","NEWT:1336795","NEWT:644042","NEWT:150808","NEWT:709964","NEWT:1182590","NEWT:3712","NEWT:3711","NEWT:156974","NEWT:3714","NCBITaxon:37162","NEWT:270643","NEWT:3715","NEWT:1538","NEWT:2065263","NEWT:199225","NEWT:1549675","NEWT:177437","NEWT:717966","NEWT:9014","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:29022","NEWT:44664","NEWT:43346","NEWT:3702","NEWT:1245466","NEWT:3704","NEWT:244366","NEWT:96795","NEWT:1246791","NEWT:176597","NEWT:118694","NEWT:2850","NEWT:118691","NEWT:9005","NEWT:34871","NEWT:9006","NEWT:33548","NEWT:52299","NEWT:1883426","NEWT:8165","NEWT:96794","NEWT:10401","NEWT:10886","NEWT:3708","NEWT:332648","NEWT:44670","NEWT:926571","NEWT:536231","NEWT:376219","NEWT:219813","NEWT:1510","NEWT:1955249","NEWT:215451","NEWT:460519","NEWT:1515","NEWT:572307","NEWT:1432138","NEWT:2714896","NEWT:1424507","NEWT:1194599","NEWT:272844","NEWT:1348799","NEWT:1608994","NEWT:33995","NEWT:55784","NEWT:1303443","NEWT:9483","NEWT:485","NEWT:329032","NEWT:56636","NEWT:2709072","NEWT:2499525","NEWT:55307","NEWT:1122981","NEWT:2853422","NEWT:1122984","NEWT:487","NEWT:1538553","NEWT:1679718","NEWT:480","NEWT:67767","NEWT:45972","NEWT:2830","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:219809","NEWT:219808","NEWT:119562","NEWT:1883446","NEWT:31345","NEWT:1233435","NEWT:118223","NEWT:11757","NEWT:141469","NEWT:987059","NEWT:109760","NEWT:29031","NEWT:317047","NEWT:1509","NEWT:491","NEWT:460523","NEWT:45954","NEWT:1578","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:291146","NEWT:499175","NEWT:109779","NEWT:3745","NEWT:1715989","NCBITaxon:4751","NEWT:3747","NEWT:278161","NEWT:1116234","NEWT:415978","NEWT:235455","NEWT:1255228","NEWT:410289","NEWT:191701","NEWT:373153","NEWT:1640278","NEWT:9054","NEWT:1092586","NEWT:472","NEWT:940825","NEWT:29459","NEWT:529507","NEWT:1071661","NEWT:1380096","NEWT:28128","NEWT:470","NEWT:41122","NEWT:5911","NEWT:1328300","NEWT:748693","NCBITaxon:50557","NEWT:39251","NEWT:29491","NEWT:911022","NEWT:101841","NEWT:446","NEWT:153481","NEWT:2014887","NEWT:717989","NEWT:33952","NEWT:1255232","NEWT:11723","NEWT:445","NEWT:153009","NEWT:1004879","NEWT:261756","NEWT:166314","NEWT:16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