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. 2014 Jan 14;34(1):e00089. doi: 10.1042/BSR20130091

Figure 5. Involvement of p38 kinase in TGFβ1/ROS-dependent expression of αSMA.

Figure 5

(A) Subconfluent HDFs were preincubated with MAPK inhibitors U0126, SP600125 or SB202190 before treatment with rTGFβ1. Expression of αSMA was detected by Western blots. The densitometric analysis describes protein expression as fold increase over control, which was set at 1.0. The data represent the mean±S.E.M. of three independent experiments. (B) Subconfluent HDF were preincubated for 1 h with SB202190 (10 μM) in the serum-free medium and then rTGFβ1 (10 ng/ml) treated for 24 h. αSMA mRNA levels were analysed by real-time RT-PCR. Data are given as means of three independent experiments±S.E.M. (C) Subconfluent HDFs were either mock-treated, treated with rTGFβ1 (10 ng/ml) for 48 h or incubated with SB 202190 for 48 h or starting 4, 8 and 16 h after rTGFβ1 treatment. The level of αSMA protein was determined by Western blot. α-tubulin was used as loading control. Three independent experiments were performed. (D) Subconfluent HDFs were either mock-treated or pretreated for 1 h with apocynin (1 mM) before addition of rTGFβ1 (10 ng/ml). TGFβ1 and apocynin were present for an additional 12 h. Anisomycin (0.5 μg/ml) was used as technical control and incubated for 20 min. The level of phospho-p38 MAPK protein was determined by Western blot. α-tubulin was used as loading control. Two independent experiments were performed.