Fig. 6
E2F1 induces the expression of EZH2 in GC B cells. a Expression level in FPKM of E2F genes in naive B and GC B cells from 4 human tonsils. Horizontal black lines represent mean. t test, ***P < 0.001. b RT-qPCR of E2f in GCBs (FAS + GL7 + B220 + ) and naive B cells (FAS-GL7-IgD + B220 + ) sorted from 7 WT mice. Horizontal black lines represent mean fold change mRNA levels normalized to Hprt1. t test, ***P < 0.001. c Immunoblotting of whole cell lysates from naive B and GC B cells from two human tonsil samples. Actin was used as loading control. The western blot shown is representative of a total of five tonsils analyzed. See Supplementary Fig. 6B for uncropped scans of the western blots. d Representative immunoblotting of whole cell lysates from OCI-Ly1 and OCI-Ly7 cells expressing shRNAs against E2F1, E2F2 or control. GAPDH was used as loading control. The experiment was repeated two more times with similar results. See Supplementary Fig. 6C for uncropped scans of the western blots. e Schematic representation of the genomic region corresponding to the promoter of EZH2. The black boxes represent a region of putative E2F binding sites. Two pairs of primers were design to flank the putative E2F binding sites. f E2F1 and control IgG qChIP was performed in the indicated cell lines using the primers described in e. As negative control qPCR was performed using primers for two regions in chromosomes 6 and 7 where no enrichment of transcription factors was found by ChIP-seq read density in OCI-Ly1 and OCI-Ly7 cells. As positive control qPCR was performed using primers flanking an E2F binding site at the promoter of CDK1. Fold enrichment was normalized to the input. Values are shown as mean of technical triplicates ± SEM. The experiment shown is representative of three independent ChIPs performed with OCI-Ly1 and OCI-Ly7, and two with WSU-DLCL2
