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. Author manuscript; available in PMC: 2017 Aug 1.
Published in final edited form as: Biochim Biophys Acta. 2016 May 20;1859(8):1043–1055. doi: 10.1016/j.bbagrm.2016.05.009

Figure 3. Knockdown of Ezh2 results in the expression of Runx2/p57 in immature primary hippocampal cells.

Figure 3

Hippocampal cells were infected at 3DIV with a lentivirus carrying a shRNA against Ezh2 expression. (A): Total RNA samples were obtained 96h later (7DIV) to confirm the Ezh2 silencing by qRT-PCR (left panel). Results were normalized against GAPDH and expressed as fold change with respect to cells infected with a lentivirus packed with an empty vector. Ezh2 knockdown was also confirmed by Western blot using specific antibodies (right panel). Detection of RNA Polymerase II with a specific antibody was used to control for equal protein loading. (B-E): ChIP experiments were performed on samples using specific antibodies against total H3 (B), H3K27me3 (C), H3K4me3 (D) and H3K27ac (E). Results are expressed as Input (%) ± SD of at least three independent experiments. Normal IgG was used as specificity control. Primers for PSD95 promoter region were used as positive controls. (F): Changes in mRNA expression of osteoblastic genes were determined by qRT-PCR. *: P < 0.05, **: P < 0.01, ns: non-significant differences with respect to the corresponding control (empty vector) value (Student's t-test).