Fig. 7
E2F1 is required for GC formation in an EZH2 dependent manner. a–f E2f1 −/− and control WT mice were immunized with SRBC to induce GC formation and were killed 10 days later. a Average of percentage of B cells (B220 + ) within live splenocytes (DAPI−) of each group of mice (n = 9 per group). b 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). c Average of percentage of GC B populations of each group quantified by flow cytometry as in b (n = 9 per group). d Formalin fixed paraffin embedded splenic tissue was stained for PNA, Ki67, EZH2, phospho Rb Ser780 and B220. One representative picture of 5 spleens analyzed per group is shown. e, f Quantification of PNA staining from d (n = 5 spleens per group). e “#GC/spleen section” is the count of all GC per spleen section. f “GC area/total spleen area” is the quantified area of each individual GC divided by the total area of the spleen section. Results shown in a–f are representative of a total of three independent experiments performed with different cohorts of mice. g Bone marrow transplantation was performed using E2f1 −/− and WT donor mice. Each recipient group consisted of 8 mice. BM, bone marrow. h Gating strategy used to analyze the GFP positive GC B cells from transplanted mice. Representative flow cytometry plots showing the gating on GC B cells (GL7 + FAS + ) within live GFP B cells (B220 + DAPI-GFP + ) per mouse group. i Average of percentage of GFP positive GC B cells in each transplant group (n = 8) quantified by flow cytometry as shown in h. Values in a, c, e, f, i are shown as mean ± SEM. t test, **P < 0.01, ***P < 0.001. Results shown in a–f are representative of a total of four independent experiments
