Fig. 2: IgG4-RD patients have immunocompromise corresponding to reduced USP25 expression.

A Enriched KEGG pathway interaction networks in B cells from HCs and patients (n = 5). B, C Integration analysis results of IgG4-RD and HCs, showing the B cell clusters (B) and status (C) (n = 3). D Dot plot showing the expression of selected canonical cell markers in the 11 B cell subsets in patients and HCs. E The expression level of USP25 on B cell subsets in IgG4-RD and HCs showed by UMAP separately (n = 3). F Relative mRNA level of USP25 was measured in total B cells (n = 5), memory B cells (n = 3) and naive B cells (n = 3) from HCs and patients after sorting. G, H Purified B cells of patients and HCs were stimulated, fixed, permeabilized, and stained for USP25 (scale bar = 2.5 μm). The correlation coefficient between BCR and USP25 was quantified using more than 47–101 individual cells (H). I Western blots of USP25 in purified B cells from HCs and patients. J Western blots of USP25, LYN, pSYK, SYK, pBLNK and BLNK in PBMCs. K Western blots of USP25 levels in purified B cells from WT and Usp25 KO mice. L Western blots of LYN, pSYK, SYK, pBLNK, BLNK, pSHIP, SHIP, pWASP, and WASP in B cells of WT and Usp25 KO. M Immunoassay of lysates of WT cells treated with 10 µg/ml sAg for 5 min, followed by immunoprecipitation with LYN and immunoblot analysis of USP25. N Immunoassay of lysates of WT and Usp25 KO B cells treated in the same way as in (M), followed by immunoprecipitation with LYN and immunoblot analysis with Ubiquitin. O Immunoblot showing the levels of USP25, LYN, pAKT, pFOXO1 and pS6. PBMCs were treated with 25 μM of MG132 at 37 °C for 30 min before incubating in the same way as in (Fig. 1J) for 5 min. All images were representative images from 3 independent experiments. Data points in (E, F, and H) are represented as Mean ± SED. Statistical significance was based on two-tailed unpaired Student’s t-test and wilcoxon rank sum test. Relevant p values are given in the graph. *P < 0.05; **P < 0.01; ***P < 0.001. Source data are provided as a Source Data file.