Skip to main content
. Author manuscript; available in PMC: 2015 Dec 1.
Published in final edited form as: J Bone Miner Res. 2014 Dec;29(12):2676–2687. doi: 10.1002/jbmr.2296

Fig. 8.

Fig. 8

GATA4 regulates the TGFβ and BMP pathways in osteoblasts. (A) Calvarial osteoblasts were infected with lentivirus expressing either shGFP or shGATA4. Cells were then differentiated for 2 wk and RNA was obtained. qPCR was performed for the indicated genes and normalized to actin mRNA. (B) Calvarial osteoblasts were infected with lentivirus expressing either shGFP or shGATA4. Cells were then differentiated for 2 wk and then treated with E2 for 24 hours. Protein was obtained and immunoblots for pSMAD1/5/8, SMAD5, GATA4, and actin were performed. (C) RNA was obtained as in A, and qPCR was performed for the indicated genes and normalized to actin mRNA. (D) Protein was obtained as in B, and immunoblots for pSMAD3, pSMAD2, GATA4, and actin were performed. (E) IHC with an antibody to pSMAD1/5/8 was performed on femurs from P0 WT (Fl/Fl) and cKO (Fl/Fl; Cre+) mice. (F) Immunofluorescence with an antibody to pSMAD1/5/8 was performed on femurs from P0 WT (Fl/Fl) and cKO (Fl/Fl; Cre+) mice. *p < 0.05; **p < 0.001. cKO = conditional knockout; h.ch = hypertrophic chondrocytes; tr = trabecular bone; IHC = immunohistochemistry.