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. Author manuscript; available in PMC: 2014 Jan 15.
Published in final edited form as: Cancer Res. 2012 Nov 7;73(2):695–705. doi: 10.1158/0008-5472.CAN-12-2522

Figure 4. Ezh2 alone can repress MYC and Myc-repressed genes.

Figure 4

(A) Ezh2 and Ezh2 mutants were stably expressed in myc−/− fibroblasts.

(B) Expression of the MYC promoter, GADD45 and PDGFRb were assessed by RT-PCR.

(C) ChIP for H3K27me3 levels at the MYC, GADD45 and PDGFRb promoters in the same lines from B.

(D) ChIP for H3K4me3 levels at the MYC, GADD45 and PDGFR promoters in the same lines from B.

(E) Immunoblot analysis of total cell lysates from myc+/+ and myc−/− samples expressing empty Vector (Vec), exogenous Myc (Myc) or Ezh2 S-A. Myc and Ezh2 increase global levels of H3K27me3.

(F) Heat map of repressed genes from microarray data. Blue (downregulated) and yellow (upregulated). The bracket marks genes that are repressed by both Myc and Ezh2S21A. Venn diagram demonstrates the overlap of genes repressed by 2-fold or more in both samples.

(G) Soft agar transformation assay with IMEC cells expressing empty vector (Vec) or exogenous Myc (Myc) upon depletion of Ezh2 with RNAi. Graph represents mean number of colonies that were larger than 10µM per 100 colonies counted (48). Data presented is average of three independent replicates and error bars indicate S.D.