Supplementary Figure 4: IRF4hi and IRF8hi activated B cells manifest distinct gene-expression states. | Nature Immunology

Supplementary Figure 4: IRF4hi and IRF8hi activated B cells manifest distinct gene-expression states.

From: Regulation of bifurcating B cell trajectories by mutual antagonism between transcription factors IRF4 and IRF8

Supplementary Figure 4

(a) Naïve splenic B cells were isolated and stimulated with LPS or anti-CD40 plus IL-2, IL-5 and IL-4 as described in Methods. B cells were harvested and analyzed for IgG3 or IgG1 CSR at 72 h post activation.

(b) Flow sorting gates use to enrich IRF4hi (IgMhiCD138hi, indicated as IRF4hi) or IRF8hi (IgMloCD138lo, indicated as IRF8hi) cells (left panel). IRF4 and IRF8 protein amounts in IRF4hi and IRF8hi B cells analyzed by western blotting (right panel).

(c) Irf4 and Irf8 transcripts in indicated cells were measured by Q-PCR. Pou2f1 transcripts were used for normalization. Data are from three independent experiments (mean±SEM, *P < 0.01, **P < 0.001).

(d) Selected GO functional categories for differentially expressed genes in IRF4hi and IRF8hi cells are displayed with their odds ratios.

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