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. 2022 Sep 13;219(10):e20220903. doi: 10.1084/jem.20220903

Figure 1.

Figure 1.

Glucocorticoids induce IgE switching in B cells. (A) Experimental setup and representative plots for assessing serum starvation effects on ex vivo B cell class switching. CD19+ splenic B cells were cultured with IL-4 and aCD40 in control complete media (Cont, gray) or with SF conditions (red). Numbers indicate the percentage of IgE+ IgG1− or IgG1+ IgE− B cells within a live B220+ population, determined by flow cytometry (n = 11–12 mice/group, compiled from three independent experiments). (B) Percentage of IgE+ IgG1− B cells (within live B220+) from B cells cultured in Cont media (gray), SF conditions (red), SF conditions with DEX (teal), or SF conditions with Cort (orange), as described in A (n = 6–9 mice/group, compiled from three independent experiments). (C) Experimental setup and representative plots for assessing glucocorticoid effects on ex vivo B cell switching. B cells cultured in Cont media were treated in two 6-h windows with vehicle control (Cont, gray) or with DEX or Cort (teal). Numbers indicate the percentage of IgE+ IgG1− B cells within a live B220+ population by flow cytometry (n = 6 mice/group, compiled from two and representative of five independent experiments). (D) Relative RT-qPCR expression of mature Ighe, Ighg1, or Ighm transcripts (top) and upstream εGLTs, γ1GLTs, or μGLTs (bottom) for B cells cultured with Cont (gray) or DEX (red). Expression at 96 h was normalized by Rpl13a (n = 3 mice/group). (E) The percentage of IgE+ IgG1− B cells (within live B220+) from B cells cultured with Cont (gray) or RU-486 (red), as described in C (n = 3 mice/group, representative of two independent experiments). (F) Percentage of IgE+ IgG1− B cells (within live B220+) from Cont (gray) or DEX-treated (red) Cd19-Cre+/− GR Het vs. cKO B cells, as described in C (n = 6–7 mice/group, compiled from three independent experiments). (G) Percentage of IgE+ IgG1− B cells (within live B220+) from Cont (gray) or DEX-treated (red) Mb1-Cre+/− GR Het vs. cKO B cells (n = 3 mice/group). Error bars in A–G represent SEM. P values <0.05 were considered significant (*, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; ns [or unlabeled], not significant). Ratio paired Student’s t test was used for A–G.