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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 6. Overexpression of Wdr5 results in the expression of Runx2/p57 in primary hippocampal cells.

Figure 6

2DIV rat calvaria-derived osteoblasts (A) and 3DIV hippocampal cells were infected or transfected, respectively, with a vector coding for the COMPASS subunit Wdr5 (B-J). (A, B): Total RNA samples were obtained 96h later to confirm Wdr5 mRNA overexpression by qRT-PCR (left panels). Results were normalized against GAPDH and expressed as fold change with respect to cells transfected with an empty vector. Wdr5 overexpression was also confirmed by Western blot using specific antibodies (right panels). TFIIB protein detection was used to control for equal loading. (C-E, G-J): ChIP experiments were performed on samples using specific antibodies against Wdr5 (C), H3K4me3 (D), H3K27me3 (E), UTX (G), JmjD3 (H, I), and Ezh2 (J). Results are expressed as Input (%) ± SD of at least three independent experiments. Normal IgG was used for specificity control. (A [right panel] and F): Changes in mRNA expression of Runx2/p57 were determined by qRT-PCR. *: P < 0.05, **: P <0.01, ***: P<0.001, ns: non-significant differences with respect to the corresponding control (pCDH) value (Student's t-test). For panel H, **: P<0.01, with respect to E18 (ANOVA).