Skip to main content
. 2021 Dec 1;12:756131. doi: 10.3389/fphar.2021.756131

FIGURE 4.

FIGURE 4

Inhibiting Sirt2 activity and expression downregulates the increased Smad2/3 phosphorylation induced by TGF-β1. (A) Protein expression of p-Smad2/Smad2 and p-Smad3/Smad3 by Western blot in MRC-5 cells pretreated with 2 ng/ml TGF-β1 for 24 h and then 10 μM AGK2 or DMSO for 24 h in the presence of TGF-β1. (B) Densitometric analyses of the Western blot in (A). (C) Protein expression of p-Smad2/Smad2 and p-Smad3/Smad3 by Western blot in MRC-5 cells transfected with NC siRNA or Sirt2 siRNA for 24 h in the absence or presence of 2 ng/ml TGF-β1. (D) Densitometric analyses of the Western blot in (C). (E) MRC-5 cells were treated with TGF-β1 at 2 and 5 ng/ml for 24 h, and total protein was co-immunoprecipitated with anti-Sirt2 antibody or IgG and immunoblotted with Smad3 antibody. GAPDH was used as a loading control. The bars indicated mean ± SD of three separate experiments. NC, negative control; siRNA, small interfering RNA; DMSO, dimethyl sulfoxide. *p <0.05, ***p <0.001.