Figure 2.

lnc-TSI Inhibited TGF-β1-Induced Smad3 Phosphorylation and nu. Translocation of the Smads Complex in Caki-1 Cells

(A) Western blot showed that knocking out lnc-TSI promoted Smad3, but not Smad2, phosphorylation in Caki-1 cells in the presence or absence of exogenous TGF-β1 (A₁). The data analysis results are shown in (A₂) and (A₃). (B) Western blot demonstrated that the overexpression of lnc-TSI inhibited Smad3, but not Smad2, phosphorylation in Caki-1 cells in the presence or absence of exogenous 10 ng/mL of TGF-β1 (B₁). The data analysis results are shown in (B₂) and (B₃). (C) Immunofluorescence confocal images showed that knocking out lnc-TSI enhanced Smad3 nu. translocation in Caki-1 cells while overexpressing lnc-TSI inhibited Smad3 nu. translocation in the presence or absence of exogenous 10 ng/mL of TGF-β1 for 1 h (C₁). The quantitative data of positive nu. Smad3 staining cells are shown in (C₂). (D) Western blot in nucleus and cyto. of Caki-1 cells demonstrated that knockout of lnc-TSI promoted the nu. translocation of Smads in Caki-1 cells incubated with or without exogenous TGF-β1 (D₁). β-Actin and lamin A/C were separately applied as the loading control for the cyto. or nucleus. The data analysis results are shown in (D₂), (D₃), and (D₄). (E) Western blot demonstrated that the overexpression of lnc-TSI inhibited the nu. translocation of Smads in Caki-1 cells incubated with or without TGF-β1 (E₁). The data analysis results are shown in (E₂), (E₃), and (E₄). Data are expressed as means ± SD of three independent experiments. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.