{"database":"Pride","file_versions":[{"headers":{"Content-Type":["application/json"]},"body":{"files":{"Xlsx":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD069867/20241118_SJMS_Eclipse2_FAIMS_Stanford_Profiling_9samples_protein_peptide.xlsx"],"Txt":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD069867/checksum.txt"],"Raw":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD069867/20241126_SJMS_Eclipse2_FAIMS_Stanford_Profiling_9samples_3.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD069867/20241126_SJMS_Eclipse2_FAIMS_Stanford_Profiling_9samples_2.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD069867/20241126_SJMS_Eclipse2_FAIMS_Stanford_Profiling_9samples_4.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD069867/20241126_SJMS_Eclipse2_FAIMS_Stanford_Profiling_9samples_1.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD069867/20241126_SJMS_Eclipse2_FAIMS_Stanford_Profiling_9samples_6.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD069867/20241126_SJMS_Eclipse2_FAIMS_Stanford_Profiling_9samples_5.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD069867/20241126_SJMS_Eclipse2_FAIMS_Stanford_Profiling_9samples_9.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD069867/20241126_SJMS_Eclipse2_FAIMS_Stanford_Profiling_9samples_7.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD069867/20241126_SJMS_Eclipse2_FAIMS_Stanford_Profiling_9samples_8.raw"]},"type":"primary"},"statusCode":"OK","statusCodeValue":200}],"scores":null,"additional":{"labhead_mail":["dokwan@stanford.edu"],"submitter":["Viktoria Szeifert"],"technology_type":["Mass Spectrometry","Bottom-up proteomics"],"software":["Not available"],"submitter_keywords":["Ev","Exosome","Pmn ev ms","Extracellular vesicles","Neutrophil"],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD069867"],"tissue":["Neutrophil","Blood"],"sample_protocol":["Nanoflow liquid chromatography–tandem mass spectrometry was performed on EVs isolated from three biological replicates. The dry EV pellets were resuspended in 10 μL sterile distilled water (nuclease-free), merged for bulk analysis, snap frozen, and stored at -80˘C until processing within 1 week. The samples were first lysed in 5% SDS, 9M urea, and 50mM TEAB buffer, sonicated, and centrifuged. Protein concentration of the supernatant was then measured by BCA assay, followed by reduction in 10mM DTT and alkylation in 20mM IAM. Subsequently, the samples were subjected to clean up and Lys-C/trypsin digestion in S-Trap micro columns (ProtiFi) according to the manufacturer’s protocol. Finally, the digested peptides were eluted from the plate and dried by a SpeedVac vacuum concentrator (Thermo Fisher). Samples were analyzed using an Ultimate 3000 nanoflow HPLC (Thermo Fisher Scientific) coupled to an Orbitrap Eclipse™ Tribrid™ mass spectrometer (Thermo Fisher Scientific) with a Nanospray Flex™ ion source. Peptides were separated on a heated Aurora Ultimate nano column (250 mm × 75 µm ID, 1.7 µm C18, IonOpticks) over a 120-min water/acetonitrile gradient at 0.3 µL/min. MS data were searched against the reviewed Homo sapiens UniProt database using Proteome Discoverer 2.5 (Sequest algorithm)."],"repository":["Pride"],"quantification_method":[""],"modification":[""],"data_protocol":["For downstream analysis, protein-level abundances were log₂-transformed as log₂ (intensity + 1) to stabilize variance. Contaminants and unchecked identifications were removed. Proteins were ranked by summed abundance across conditions, and the top 100 most abundant were visualized as row-scaled heatmaps. In addition, a curated subset of neutrophil granule proteins was analyzed separately to highlight their abundance patterns across inhibitor treatments. Heatmaps were generated in R using pheatmap (version 1.0.12) with case–control group annotations."],"omics_type":["Proteomics"],"labhead":["Derick Okwan-Doudu"],"instrument_platform":[""],"labhead_affiliation":["Assistant Professor"],"submission_type":["PARTIAL"],"species":["Homo Sapiens (human)"],"submitter_mail":["szeivik@stanford.edu"],"publication":["42000854 Szeifert V, Szeifert A, Arnold WR, Terasaki A, Bhatnagar K, Okwan-Duodu D. Comparative analysis of extracellular vesicles released from resting neutrophils under distinct pharmacological inhibition. Sci Rep. 2026 16(1):18092 10.1038/s41598-026-48570-8"],"submitter_affiliation":["Postdoctroal Fellow"],"submitter_country":["United States"],"pubmed_abstract":["Cells in resting conditions spontaneously release extracellular vesicles (EVs). Comparative studies for pharmacological EV inhibition, targeting multiple biogenesis elements within the same cell type, remain scarce. Spontaneously released neutrophil-derived EVs are important mediators of immunity that exhibit anti-inflammatory properties. Here, we present a systematic investigation of seven pharmacologic inhibitors (calpeptin, D-pantethine, Y27632, cytochalasin D, GW4869, R5421, and Nexinhib20) on the quantitative and qualitative properties of the selected EV population (16,100 g pellet obtained by filtration, size‑exclusion chromatography, and differential centrifugation) derived from resting neutrophils. We measured EV numbers, size distribution, total lipid, protein, and RNA content. We also analyzed EV morphology by cryogenic electron microscopy and EV cargo composition qualitatively by mass spectrometry and RNA sequencing. The results revealed differential profiles of proteins, mRNAs, and miRNAs depending on the inhibitor applied, as well as changes in the physical characteristics of the EVs. Inhibition of a single biogenesis pathway often led to compensatory upregulation of alternative vesicle formation mechanisms, as reflected in preserved EV numbers but altered cargo composition. These findings underscore the necessity of carefully selecting and reporting EV-inhibiting pharmacologic agents, as each compound uniquely shapes both the quantity and biological cargo of released EVs, and may have physiological consequences."],"pubmed_title":["Comparative analysis of extracellular vesicles released from resting neutrophils under distinct pharmacological inhibition."],"pubmed_authors":["Szeifert Viktória V, Szeifert Alexa A, Arnold William Randall WR, Terasaki Azusa A, Bhatnagar Keshav K, Okwan-Duodu Derick D"],"additional_accession":[]},"is_claimable":false,"name":"One way or another: Differential impact of pharmacologic inhibitors on extracellular vesicles derived from resting neutrophils","description":"We performed a systematic investigation of seven pharmacologic inhibitors (calpeptin, D-pantethine, Y27632, cytochalasin D, GW4869, R5421, and Nexinhib20) on quantitative and qualitative EV properties derived from resting neutrophils. We measured EV numbers, size distribution, total lipid, protein, and RNA content. We also analyzed the EV morphology by cryo-EM and EV cargo qualitatively by mass spectrometry and RNA sequencing.","dates":{"publication":"2026-06-15","submission":"2025-10-24"},"accession":"PXD069867","cross_references":{"TAXONOMY":["NEWT:330879","NEWT:377960","NEWT:2042546","NEWT:259447","NEWT:295546","NCBITaxon:2719036","NCBITaxon:1280","NEWT:112503","NEWT:1129","NEWT:309807","NEWT:59753","NEWT:89184","NEWT:309800","NEWT:281395","NEWT:1211601","NEWT:876138","NEWT:44271","NEWT:193516","NEWT:1073882","NEWT:475174","NEWT:111205","NEWT:1117","NEWT:498257","NEWT:10036","NEWT:1590","NEWT:10034","NEWT:661410","NEWT:638632","NEWT:224326","NEWT:376619","NCBITaxon:79857","NEWT:1096976","NEWT:1589","NEWT:135622","NEWT:35786","NEWT:1580","NEWT:399784","NEWT:96731","NEWT:383379","NEWT:418106","NEWT:10029","NEWT:913645","NEWT:641809","NEWT:38815","NEWT:317447","NEWT:4688","NEWT:111225","NEWT:7719","NEWT:868565","NEWT:135674","NEWT:79329","NEWT:30069","NEWT:12637","NEWT:59729","NEWT:2164133","NEWT:295105","NEWT:108061","NEWT:60711","NEWT:224308","NEWT:3347","NEWT:160621","NEWT:212790","NEWT:110368","NEWT:1310161","NEWT:77133","NEWT:145481","NEWT:1310165","NEWT:29058","NCBITaxon:79824","NEWT:1912919","NEWT:44688","NEWT:44689","NEWT:498211","NEWT:347256","NEWT:5518","NEWT:398007","NEWT:1527468","NEWT:498217","NEWT:498216","NEWT:11320","NEWT:246196","NEWT:246197","NEWT:145458","NEWT:44685","NEWT:161934","NEWT:1148","NEWT:5508","NEWT:3329","NEWT:419481","NEWT:5507","NEWT:410661","NEWT:1140","NCBITaxon:2157","NEWT:1143","NEWT:1287689","NEWT:1094343","NEWT:1462472","NEWT:1336795","NEWT:644042","NEWT:1182590","NEWT:3712","NEWT:3711","NEWT:270643","NEWT:2065263","NEWT:177437","NEWT:10418","NEWT:118698","NEWT:1616117","NEWT:118696","NEWT:34865","NEWT:52283","NEWT:284812","NEWT:8175","NEWT:43330","NEWT:1603293","NEWT:44664","NEWT:43346","NEWT:3702","NEWT:1245466","NEWT:244366","NEWT:1246791","NEWT:118694","NEWT:2850","NEWT:118691","NEWT:34871","NEWT:33548","NEWT:52299","NEWT:8165","NEWT:96794","NEWT:3708","NEWT:332648","NEWT:44670","NEWT:536231","NEWT:376219","NEWT:219813","NEWT:1510","NEWT:460519","NEWT:1515","NEWT:572307","NEWT:1432138","NEWT:1424507","NEWT:1194599","NEWT:272844","NEWT:1303443","NEWT:9483","NEWT:485","NEWT:56636","NEWT:2709072","NEWT:2853422","NEWT:1679718","NEWT:480","NEWT:67767","NEWT:46835","NEWT:109757","NEWT:582580","NEWT:1185650","NEWT:216778","NEWT:294607","NEWT:1502","NEWT:128017","NEWT:376686","NEWT:95486","NEWT:1883446","NEWT:1233435","NEWT:109760","NEWT:29031","NEWT:317047","NEWT:460523","NEWT:235443","NEWT:1757312","NEWT:85694","NEWT:108458","NEWT:5936","NEWT:320637","NEWT:1078283","NEWT:3750","NEWT:983964","NEWT:11706","NEWT:32644","NEWT:527796","NEWT:499175","NEWT:109779","NEWT:3745","NEWT:1715989","NCBITaxon:4751","NEWT:3747","NEWT:1116234","NEWT:1255228","NEWT:410289","NEWT:373153","NEWT:472","NEWT:940825","NEWT:529507","NEWT:1071661","NEWT:470","NEWT:5911","NCBITaxon:50557","NEWT:39251","NEWT:29491","NEWT:101841","NEWT:446","NEWT:153481","NEWT:2014887","NEWT:33952","NEWT:445","NEWT:153009","NEWT:261756","NEWT:166313","NEWT:63366","NEWT:63367","NEWT:215402","NEWT:1547","NEWT:906914","NEWT:9031","NEWT:27292","NEWT:1050722","NEWT:108931","NEWT:1293497","NEWT:1055524","NEWT:150475","NEWT:267872","NEWT:172269","NEWT:9534","NEWT:5180","NEWT:256737","NEWT:9541","NEWT:8694","NEWT:33936","NEWT:8692","NEWT:2903","NEWT:185579","NEWT:13076","NEWT:1006581","NEWT:8673","NEWT:33940","NEWT:550","NEWT:554","NEWT:451516","NEWT:552","NEWT:1325291","NEWT:36185","NEWT:1054211","NEWT:1225786","NEWT:51338","NEWT:575412","NEWT:28112","NEWT:6493","NEWT:6494","NEWT:6491","NEWT:507601","NEWT:520","NEWT:186441","NEWT:643680","NEWT:214092","NCBITaxon:6157","NEWT:13095","NEWT:162425","NEWT:104105","NEWT:216257","NEWT:9986","NEWT:8654","NEWT:8658","NEWT:1268063","NEWT:8657","NEWT:9983","NEWT:8655","NEWT:5147","NEWT:28108","NEWT:28104","NEWT:407821","NEWT:568703","NCBITaxon:2","NEWT:568708","NEWT:986","NEWT:52641","NEWT:28532","NEWT:353152","NEWT:298176","NEWT:40674","NEWT:1194669","NEWT:51329","NEWT:443906","NEWT:519","NEWT:2510939","NEWT:7394","NEWT:6063","NEWT:1328388","NEWT:1548728","NEWT:667127","NEWT:9557","NEWT:377586","NEWT:300641","NEWT:39655","NEWT:9554","NEWT:38323","NEWT:256318","NEWT:206411","NCBITaxon:6191","NEWT:229533","NEWT:214053","NEWT:80863","NEWT:90675","NEWT:52638","NEWT:57075","NEWT:1666905","NEWT:8697","NEWT:8695","NEWT:884204","NEWT:1123869","NEWT:9544","NEWT:9545","NEWT:979","NEWT:7370","NEWT:83906","NEWT:99488","NEWT:1134506","NEWT:255470","NEWT:38783","NEWT:6426","NEWT:33090","NEWT:9935","NEWT:287889","NEWT:305959","NEWT:13489","NEWT:29829","NEWT:92867","NEWT:92866","NCBITaxon:3055","NEWT:51750","NEWT:202950","NEWT:295027","NCBITaxon:11320","NEWT:632957","NEWT:9925","NCBITaxon:9606","NEWT:90690","NEWT:1436183","NEWT:300273","NEWT:4232","NEWT:416348","NEWT:11298","NEWT:4236","NEWT:7742","NEWT:196627","NEWT:200308","NEWT:200302","NEWT:870435","NEWT:9913","NEWT:9915","NEWT:105841","NEWT:2666255","NEWT:999810","NCBITaxon:5693","NEWT:28995","NEWT:1392998","NEWT:380394","NEWT:226900","NEWT:1266738","NEW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