Fig. 3: USP25 deficiency causes the increase of IgG1.

A–C Flow cytometry analysis of IgG1 in B cells, PC and PBC from WT and Usp25 KO mice after stimulating with 8 ng/ml IL4 plus 10 μg/ml of CD40 for 120 h and 10 μg/ml of LPS for 72 h and shown are representative dot plots (A). The percentages of B cells (B), PC and PBC (C) were analyzed from WT and Usp25 KO mice (n = 6). D The level of IgG1 in PC and PBCs culture supernatant of WT and Usp25 KO mice was quantified by ELISA (n = 4). E B cells of Usp25 KO and WT mice were cultured with LPS (10 μg /mL). Total RNA was isolated after 2 days of culture and the GLTγ1 and GAPDH levels were measured by RT-PCR (n = 3). F-H Flow cytometry analysis of IgG1 in PBC (G) and PC (H) from Usp25 KO and WT mice after LCMV infection (n = 5). Shown are representative dot plots (F). I Quantification of IgG1 antibody levels in the serum of WT and Usp25 KO mice after infection with LCMV by ELISA (n = 5). J The OD of NP-IgG1 after immunized mice was quantified using ELISA (n = 4). K–M Flow cytometry analysis of IgG1 in B cells (L), PC and PBC (M) from Mb1^cre+/-Btk^fl/+ and Mb1^cre+/-Btk^fl/fl mice after stimulating in the same way as in (3 A) (n = 4). Shown are representative dot plots (K). N The level of IgG1 in PC and PBC culture supernatant of Mb1^cre+/-Btk^fl/+ and Mb1^cre+/-Btk^fl/fl mice was quantified by ELISA (n = 5). O The GLTγ1 level was measured in the same way as in (3E) in Mb1^cre+/-Btk^fl/+ and Mb1^cre+/-Btk^fl/fl mice (n = 3). P IgG1 antibody levels in the serum of Mb1^cre+/-Btk^fl/+ and Mb1^cre+/-Btk^fl/fl mice were quantified by ELISA (n = 5). Data points in (B, C, D, E, G, I, J, L, M, N, and O) are represented as Mean ± SED. Statistical significance was based on two-tailed unpaired Student’s t-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.