SRF Antibody - #AF6160

Figure 5. KIF3A knockdown increases α-SMA-positive myofibroblasts among SiO2-activated MRC-5 fibroblasts. (A) Western blot showing the effects of NC-siRNA and KIF3A-siRNA on expression of KIF3A, Ac-α-Tub, and ARL13B proteins in MRC-5 fibroblasts. GAPDH was used as a loading control (n=3). (B) Densitometric analyses of KIF3A, Ac-α-Tub, and ARL13B protein expression in MRC-5 fibroblasts. *P<0.05; **P<0.01. Data are the mean±SD. Statistical analysis was performed using one-way ANOVA and SPSS 20.0. (C) IF assay showing primary cilia in MRC-5 fibroblasts treated with NC-siRNA or KIF3A-siRNA. Primary cilia were labelled with an anti-Ac-α-Tub antibody. Scale bar=25 and 5 μm. (D) Treatment regimen of KIF3A knockdown in MRC-5 fibroblasts. MRC-5 fibroblasts were stimulated with SiO2 or serum-free medium (n=3 per group) for 12 h, and then transfected with NC-siRNA or KIF3A-siRNA until 36 h. (E) Expression of α-SMA in MRC-5 fibroblasts measured by IF. Scale bar=100 μm. (F, G) Western blot and densitometric analyses of the effects of NC-siRNA and KIF3A-siRNA on expression of COL I, α-SMA, MRTF-A and SRF proteins in MRC-5 fibroblasts with or without SiO2 stimulation. α-Tub was used as a loading control (n=3). *P<0.05; **P<0.01. Data are the mean±SD. Statistical analysis was performed using one-way ANOVA and SPSS 20.0.

Figure 5 KIF3A knockdown increases α-SMA-positive myofibroblasts among SiO2-activated MRC-5 fibroblasts. (A) Western blot showing the effects of NC-siRNA and KIF3A-siRNA on expression of KIF3A, Ac-α-Tub, and ARL13B proteins in MRC-5 fibroblasts. GAPDH was used as a loading control (n=3). (B) Densitometric analyses of KIF3A, Ac-α-Tub, and ARL13B protein expression in MRC-5 fibroblasts. *P<0.05; **P<0.01. Data are the mean±SD. Statistical analysis was performed using one-way ANOVA and SPSS 20.0. (C) IF assay showing primary cilia in MRC-5 fibroblasts treated with NC-siRNA or KIF3A-siRNA. Primary cilia were labelled with an anti-Ac-α-Tub antibody. Scale bar=25 and 5 μm. (D) Treatment regimen of KIF3A knockdown in MRC-5 fibroblasts. MRC-5 fibroblasts were stimulated with SiO2 or serum-free medium (n=3 per group) for 12 h, and then transfected with NC-siRNA or KIF3A-siRNA until 36 h. (E) Expression of α-SMA in MRC-5 fibroblasts measured by IF. Scale bar=100 μm. (F, G) Western blot and densitometric analyses of the effects of NC-siRNA and KIF3A-siRNA on expression of COL I, α-SMA, MRTF-A and SRF proteins in MRC-5 fibroblasts with or without SiO2 stimulation. α-Tub was used as a loading control (n=3). *P<0.05; **P<0.01. Data are the mean±SD. Statistical analysis was performed using one-way ANOVA and SPSS 20.0.

Fig. 1. Ac-SDKP promotes extension of primary cilia and inhibits EMyT in silicotic rats (A) Sirius red staining of lung tissues and the expression levels of KIF3A measured by IHC staining in rats exposed to silica, with or without Ac-SDKP intervention (scale bar = 100 µm). (B) Primary cilia in lung tissue observed by IF staining. The primary cilia were marked by ARL13B (green), and the silicitic nodules were marked by vimentin (red) (scale bar = 50 mm). (C) Protein expression levels of COL I, α-SMA, and E-cadherin in rats exposed to silica with or without Ac-SDKP. Quantification of the Western blots normalized to the loading control, α-Tub; data are presented as mean ± SD.