Fig. 2

Cdkn1a ^−/− GC rescue is linked to the histone methyltransferase function of EZH2. Cdkn1a ^+/+ and Cdkn1a ^−/− mice were immunized with SRBC, treated daily with GSK503 (150 mg/kg/day) or vehicle and were killed 10 days after immunization. a Flow cytometry plot of one representative mouse spleen per group. The gated area shows the percentage of GC B cells (GL7 + FAS + ) within live B cells (B220 + DAPI-, see Supplementary Fig. 2A for gating strategy). b Average of GC B populations of each group of mice quantified by flow cytometry as in a (n = 5 mice per group). c Formalin fixed paraffin embedded splenic tissue was stained for PNA, Ki67, EZH2, and B220. One representative picture of three spleens analyzed per group is shown. d–f Quantification of PNA staining from c (n = 3 spleens per group, see Fig. 1d–f). g, h Splenocytes from 4 WT and 4 Cdkn1a ^−/− mice were permeabilized and stained for GC B cells (GL7 + FAS + B220⁺, see Supplementary Fig. 2A for gating strategy) and H3K27me3 g and EZH2 h using fluorochrome-conjugated anti H3K27me3 and anti EZH2 antibodies, respectively. The histograms depict the mean fluorescence intensity (MFI) ± SD of H3K27me3 and EZH2 per group of mice. Values in b, d, e, f are shown as mean ± SEM. t test, *P < 0.05, **P < 0.01, ***P < 0.001. Results are representative of a total of three independent experiments performed with different cohorts of mice. See also Supplementary Fig. 3