Repsox represses translocation of Smad3 and Smad4 into nucleus resulting in attenuation of the effect of TGF-β1 on Cx43
(A) Western blot assay showing the expression of Smad3, p-Smad3, and Smad4 upon exposure to Repsox for 24 h. The results were based on three independent experiments (n=3). (B) Quantitation was performed to confirm the protein changes in (A) (n=3). **P<0.01. (C,D) Representative IF staining by CLSM showing reduced translocation of Smad3 (C) and Smad4 (D) in osteocytes after pretreatment with 50 μM Repsox for 6 h and then treated with 5 ng/ml TGF-β1 for 24 h compared with the group only treated with TGF-β1 (cytoskeleton, green; Smad3 and Smad4, red; nucleus, blue). The results were based on the three independent experiments (n=3). (E) Representative IF staining by CLSM showing that Repsox attenuated the Cx43 expression even in TGF-β1-treated osteocytes. (F) Quantitation further showing the changes in dendritic processes of osteocytes pre-incubated with Repsox after treatments of TGF-β1 (n=3). **P<0.01.