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. Author manuscript; available in PMC: 2012 Jun 15.
Published in final edited form as: Gene. 2011 Feb 16;479(1-2):16–28. doi: 10.1016/j.gene.2011.02.002

Fig. 3.

Fig. 3

Runx2 is required for efficient Wnt pathway activation in ERα activated osteoblasts. Fetal rat osteoblasts were transfected for 24 hours to express reporter plasmids TOP-Flash, the Runx sensitive reporter SXN1C, or 5XGAL4 in combination with expression plasmid encoding native ERα (A), vector pSV7d or pSV7d encoding Runx2 in antisense orientation (αSR2) (Ji et al., 2001; McCarthy, 2003; McCarthy and Centrella, 2010), empty vector M driving the expression of GAL4DBD or vector M fused to full length Runx2 (M-Runx2; center panel B) or ERα (M-ERα, right panel B) as indicated, or with expression vector encoding native ERα (C). The cells were treated with vehicle (0), 10 nM 17βE and/or 20 mM LiCl as indicated, and reporter gene enzyme activity was measured after 24 hours. 17βE significantly increased reporter gene expression through TOP-Flash, SXN1C, and 5XGAL4 in combination with M-Runx2 or M-ERα (A,B); LiCl significantly increased reporter gene expression through TOP-Flash, SXN1C, and 5XGAL4 in combination with M-ERα (A,B); 17βE and/or LiCl together significantly co-increased reporter gene expression through TOP-Flash, SXN1C, and 5XGAL4 in combination with M-Runx2 (A,B); αSR2 expression significantly suppressed TOP-Flash activity in 17βE and/or LiCl activated cells (left panel, A); (* = P <0.05).