Fig. 5: Assessment of the effects of GSK3 activation in glutaminolysis-induced IL-10 expression in B cells.

a–d B cells were isolated from blood samples of HC subjects (n = 10) and AR patients (n = 10), and immediately analyzed by western blotting. The immunoblots show the GSK3 phosphorylation status in B cells; boxplots show blot density. e–k Naive B cells were treated with the reagents in the culture as denoted below each panel. CD40L: 1 µg/ml. Glutamine: The presence of glutamine in the culture. CpG: 10 µg/ml. BPTES: 10 µM. GSK3 inhibitor (SB216763, 10 µM). The immunoblots show the GSK3 phosphorylation status in B cells (e, h); boxplots show blot density (f, g, i, j). k Boxplots show IL-10 levels in culture supernatant. The data in boxplots are presented as median (IQR). ***p < 0.001, compared with the HC group (c) or the saline group (i, j, k). Statistical methods: ANOVA + Bonferroni test (c) or Dunnett’s test (i, j, k). The data of a, c, e, h are from one experiment that represent ten independent experiments. The cell viability was greater than 99% at the end of experiments.