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. 2017 Apr;56(4):465–476. doi: 10.1165/rcmb.2016-0104OC

Figure 2.

Figure 2.

TGF-β–induced TNS1 expression requires signaling through TGF-β receptor I but not Smads. (A) Western blot of HLF lysates after stimulation with TGF-β (1 ng/ml) at indicated time points against phosphorylated (p) Smad2, pSmad3, and phosphorylated extracellular signal-regulated kinases 1 and 2 (pErk1/2). (B) HLFs were treated with 1 ng/ml TGF-β for 30 minutes in the presence of 10 μM SB531542 or vehicle control, followed by Western blotting for the indicated (phospho) proteins. (C and D) HLFs were treated with 1 ng/ml TGF-β for 24 hours in the presence of 10 μM SB531542 or vehicle control, followed by Western blotting for TNS1 (C) and subsequent densitometry (D). (E and F) HLFs were transfected with Smad2 or Smad3 small interfering (si)RNA or scrambled control followed by treatment with 1 ng/ml TGF-β for 24 hours and Western (E) blotting and densitometry (F) for the indicated proteins. Mixed-effect ANOVA (*P < 0.05; NS, not significant) was used for all statistical analyses. Data are presented as means (± SD). scram., scrambled.