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. Author manuscript; available in PMC: 2014 Oct 25.
Published in final edited form as: Biochem Biophys Res Commun. 2013 Oct 5;440(3):454–459. doi: 10.1016/j.bbrc.2013.09.128

Figure 4. Grg4 mediated activation through inhibition of Smad7.

Figure 4

A) Grg4 does not affect BMP mediated phosphorylation of Smad1/5/8. Cells were transfected with or without Grg4 and cultured with Bmp7 for 24h or a 1h pulse followed by 24h chase. Western blots indicate the levels of P-Smad1/5/8 and flag-Grg4. B) SiRNAs against Smad4 reduce levels of SMad4 mRNA. C) Smad4 knockdown does not affect EGFP reporter gene expression in response to Grg4. Western blots show levels of EGFP, flag-Grg4, P-Smad1/5/8 and tubulin loading controls. D) Quantitative RT-PCR of Smad7 mRNA in control (GFP) or Grg4 transfected cells. E) Co-transfection of Smad7 inhibits the Grg4 mediated EGFP reporter gene expression. F) Endogenous levels of Grg4 are not affected in BMP7 treated HEK293 cells.