Fig. 5: USP25 deficiency affects B cell metabolism in IgG4-RD patients through the PI3K-AKT pathway. | Nature Communications

Fig. 5: USP25 deficiency affects B cell metabolism in IgG4-RD patients through the PI3K-AKT pathway.

From: Expression of USP25 associates with fibrosis, inflammation and metabolism changes in IgG4-related disease

Fig. 5

A Western blot of pPI3K, pAKT, PI3K, AKT, PKM2, HIF-1α, c-Myc, pSTAT5 and STAT5 in PBMCs. B, C OCR (B) and ECAR (C) detection of B cells (n = 3). D The mitochondrial structure of B cells were analyzed by TEM (scale bar = 1 μm-500 nm). E–I Purified B cells were stimulated in the same way as in (Fig. 2G) except stained for Mito and ER (scale bar = 2.5 μm). The B cells were analyzed for MFI of ER (F) and Mito (G). The correlation coefficient of ER/BCR (H) and Mito/BCR was analyzed (I). J, K Purified B were pre-incubated with 10 µg/ml sAg for 24 h at 37 °C and then stained for anti-CD19 and PK Mito. Confocal analysis of the MFI of PK Mito in CD19+ cells (scale bar = 2.5 μm) (K). L Purified splenic B cells were stimulated with 10 μg/ml sAg for 24 h at 37 °C, stained for anti-B220 and mitotracker, and then analyzed for the MFI of mitotracker Green in B220 + B cells (n = 3). M PBMCs were stimulated in the same way as in (5 J) except stained for anti-CD19 and mitotracker, and then CD19 + B cells were analyzed for the MFI of mitotracker Green (n = 3). N PBMCs were stimulated, and stained for anti-CD19 and CellIROX Green, and then CD19 + B cells were analyzed for the MFI of CellIROX Green using flow cytometry (n = 3). O Cluster analysis of the differentially expressed genes in the serum of WT and Usp25 KO mice is presented in the form of a heatmap (n = 3). P Top enriched up-regulated and down-regulated KEGG pathways for differentially expressed metabolite in the serum of Usp25 KO and WT mice. Q–T The serum levels of glutamic (n = 4) and cysteine (n = 15) were detected in mice and patients respectively. All images were representative images from 3 independent experiments. Data points in (B, C, F, G, H, I, K, L, Q, R, S, and T) 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.

Back to article page