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D075581/20250628_Harim_MB6_DIA_2Th_3ms_CAC18_Col237_150trap_V3to20_G4to37B_50min_8.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD075581/20250628_Harim_FB6_DIA_2Th_3ms_CAC18_Col237_150trap_V3to20_G4to37B_50min_6.rawprimaryOK200junqu@buffalo.eduChao XueMass SpectrometryBottom-up proteomicsData-independent acquisitionTranscriptome and histologySalivary glandSalivaTuft cellhttps://www.ebi.ac.uk/pride/archive/projects/PXD075581SalivaTotal protein concentrations of saliva samples were determined via BCA assay. For each sample, 30 µg of protein in 0.5% SDS buffer was reduced with 10 mM DTT (56°C, 30 min) and alkylated with 20 mM IAA (37°C, 30 min, dark). Proteins were precipitated with chilled acetone (-20°C, 3 h), pelleted, washed with methanol, and reconstituted in 50 mM Tris-FA. Trypsin was added at a 1:20 (w/w) ratio for overnight digestion at 37°C. The reaction was quenched with 1% FA, and samples were centrifuged at 20,000 g for 30 min at 4°C to collect the peptide supernatants for LC vials.PrideNot availableRaw MS files were analyzed using DIA-NN (v1.8.1) in library-free mode against the Uniprot-SwissProt Mouse database (January 2024). Trypsin was specified as the protease (maximum one missed cleavage). Search parameters allowed up to three modifications, including cysteine carbamidomethylation, methionine oxidation, and N-terminal acetylation. Mass accuracies were set to 3 ppm (MS1) and 7 ppm (MS2), with precursor and protein FDR both strictly controlled at 1%. For quantitative downstream analysis, protein intensities were log₂-transformed, filtered for ≥70% completeness per group, and imputed using a group-aware K-nearest neighbor method (k=2). Differential abundance was evaluated via the limma R package (v4.4.2), defining significantly altered proteins as p < 0.05 and |fold change| ≥ 1.4.ProteomicsJun QuDepartment of Pharmaceutical Sciences, University at Buffalo, The State University of New York, Buffalo, NY, USAPARTIALMus Musculus (mouse)41972672 Rusiniak ME, Shimagami L, Drumond VZ, Souza MS, Castro FLAL, Xue C, Zhang M, Qu J, Chlipala GE, Maienschein-Cline M, Silva TAD, Sousa SF, Dos Santos HT. Highlights of Tuft Cells in Mouse and Human Salivary Glands. Cells. 2026 15(7):583 10.3390/cells15070583chaoxue@buffalo.eduSUNY at BuffaloUnited StatesTuft cells (TCs) are rare chemosensory epithelial cells that regulate mucosal homeostasis in multiple organs, but their role in salivary gland (SG) biology remains poorly defined. This study aimed to define TC structure in mice submandibular glands (SMGs), determine how TC loss affects gland organization and function, and evaluate whether TC abundance in human minor SGs is associated with Sjögren's disease (SjD) features. Specifically, TC ultrastructure and ductal localization were characterized in female and male C57BL/6J mouse SMGs by transmission electron microscopy and immunostaining. Wild-type and C57BL/6J-<i>Pou2f3<sup>-/-</sup></i> (TC-deficient mouse strain) SMGs were analyzed by histology and bulk RNA-seq, and salivary function was assessed by saliva flow and proteomics. Human minor SG biopsies from SjD and non-Sjögren sicca (nSjD) patients were analyzed by immunostaining and Poisson regression. In mice SMGs, TCs showed conserved ultrastructural features and localization in both sexes. TC loss was associated with marked sex-biased transcriptome remodeling, morphological disruption, and altered saliva quantity and quality. In humans, TC counts differed between nSjD and SjD groups and were associated with salivary flow, serologic status, and histopathologic features. These findings support a role for TCs in SG epithelial integrity and suggest TC abundance as a candidate biomarker of SG dysfunction.Highlights of Tuft Cells in Mouse and Human Salivary Glands.Rusiniak Michael E ME, Shimagami Lara L, Drumond Victor Zanetti VZ, Souza Mariana Silveira MS, Castro Fernanda Luiza Araujo Lima de FLAL, Xue Chao C, Zhang Ming M, Qu Jun J, Chlipala George Edward GE, Maienschein-Cline Mark M, Silva Tarcilia Aparecida da TAD, Sousa Silvia Ferreira de SF, Dos Santos Harim Tavares HTfalseHighlights of Tuft Cells in Mouse and Human Salivary GlandsTuft cells (TCs) are rare chemosensory epithelial cells that regulate mucosal homeostasis in several organs, but their role in salivary glands (SGs) remains unclear. This study investigated TC features in both mouse and human SGs. In mouse samples, transmission electron microscopy (TEM), histological analysis, bulk RNA-sequencing and proteomic analyses were performed on submandibular glands (SMGs) from female and male C57BL/6J (wild-type) and C57BL/6J-Pou2f3-/- mice, which lack TCs. Moreover, human minor SG biopsies were evaluated by immunostaining and Poisson regression to identify clinical, serological, and histopathological features correlated with TC abundance. TEM analysis confirmed conserved TC ultrastructure—apical microvilli, a tubulovesicular system, and cytospinules—in both female and male mouse SMGs. Moreover, TC loss was associated with sex-specific transcriptional remodeling (1742 differentially expressed genes in females versus 175 in males), morphological alterations and sex-specific saliva proteomic remodeling. In human minor SG biopsies, TC counts were correlated with salivary flow, serological and histopathology features. In summary, these findings suggest that TCs are regulators of epithelial integrity and potential biomarkers of SG dysfunction2026-04-272026-03-12PXD075581NEWT:6945NEWT:3555NEWT:241368NEWT:2NEWT:157546NEWT:190802NEWT:35554NEWT:150475NEWT:9417NEWT:347515NEWT:1216979NEWT:307972NEWT:32046NEWT:544496NEWT:5180NEWT:256737NEWT:115104NEWT:1081927NEWT:67825NEWT:13076NEWT:1249668NEWT:317NEWT:1736309NEWT:7227NEWT:7469NEWT:885318NEWT:4081NEWT:876138NEWT:554NEWT:98334NEWT:237561NEWT:10036NEWT:7574NEWT:1351NEWT:7215NEWT:272563NEWT:507601NCBITaxon:79857NCBITaxon:6157NEWT:95648NEWT:746360NEWT:6239NEWT:1589NEWT:470150NEWT:135622NEWT:216257NEWT:6915NEWT:9986NEWT:101510NEWT:4054NEWT:3880NEWT:8782NEWT:1000589NEWT:1902NEWT:85962NEWT:160488NEWT:28104NEWT:317447NEWT:7955NCBITaxon:2NEWT:985076NEWT:7959NEWT:2261NEWT:4565NEWT:1264690NEWT:6192NEWT:28532NCBITaxon:38727NEWT:34305NEWT:59729NCBITaxon:183674NEWT:224308NEWT:626528NEWT:139927NEWT:4558NEWT:209285NEWT:216595NEWT:243230NEWT:8355NEWT:931281NEWT:7029NEWT:1283300NEWT:334747NEWT:61235NCBITaxon:79824NCBITaxon:4563NEWT:5755NEWT:3218NEWT:5759NEWT:1736231NEWT:436486NEWT:6287NEWT:2242NEWT:300641NEWT:4784NEWT:727NEWT:9796NEWT:725NEWT:360106NEWT:260707NEWT:287NEWT:10117NEWT:10239NCBITaxon:6191NEWT:10116NEWT:1280NEWT:1836NEWT:1735272NEWT:83334NEWT:83332NEWT:29760NEWT:260704NEWT:703612NEWT:260705NEWT:80863NEWT:44685NEWT:2697049NEWT:1148NEWT:11676NEWT:55571NEWT:100226NCBITaxon:6073NEWT:4530NEWT:4896NEWT:6279NEWT:1123869NEWT:7370NEWT:83906NEWT:6282NEWT:1134506NEWT:575584NEWT:1773NEWT:38783NEWT:8727NEWT:1895NEWT:1182590NEWT:8726NEWT:10090NEWT:935293NEWT:749200NEWT:4120NEWT:5693NEWT:8724NEWT:51511NEWT:92867NEWT:8723NEWT:990346NEWT:5334NEWT:145953NEWT:257309NEWT:230741NEWT:284812NCBITaxon:10359NCBITaxon:1313NEWT:43330NEWT:242619NEWT:44544NEWT:632957NEWT:373995NEWT:544404NEWT:3702NEWT:129249NEWT:8839NEWT:4232NEWT:990119NEWT:2758385NEWT:4113NEWT:837NEWT:11298NEWT:171101NEWT:196627NEWT:408172NEWT:5691NEWT:408170NEWT:493760NEWT:260710NEWT:627025NEWT:400772NEWT:1097677NEWT:3708NEWT:106592NEWT:1117957NEWT:9913NEWT:1432138NEWT:10312NEWT:4100NEWT:1076NEWT:6763NEWT:803NEWT:8030NEWT:29722NEWT:380394NEWT:1692259NEWT:1639NEWT:188229NEWT:3818NEWT:480NEWT:4909NEWT:180066NEWT:67767NEWT:46835NEWT:135588NEWT:1843183NEWT:95486NEWT:58002NEWT:9103NEWT:4577NEWT:5664NEWT:2157NEWT:146479NEWT:10306NEWT:1911079NEWT:8022NEWT:145943NEWT:3635NEWT:5811NEWT:1480154NEWT:235443NEWT:1274414NEWT:59202NEWT:3197NEWT:9615NEWT:10299NEWT:860688NEWT:884019NEWT:169963NEWT:36329NEWT:1147787NEWT:72407NEWT:9606NEWT:367830NEWT:157295NEWT:410289NEWT:373153NEWT:915099NEWT:74940NEWT:1450511NEWT:470NEWT:84023NEWT:9838NCBITaxon:9615NEWT:1193501NEWT:3055NEWT:6326NEWT:6689NEWT:2762NEWT:5476NEWT:1174673NEWT:562NEWT:33952NEWT:1274432NEWT:1274426NEWT:1423NEWT:4932NEWT:70448NEWT:9825NEWT:1274423NEWT:3603NEWT:698936NEWT:2759NEWT:3847NEWT:39946NEWT:9823NEWT:9940NEWT:573NEWT:9031NEWT:1274420NEWT:709141972672