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. Author manuscript; available in PMC: 2007 Nov 17.
Published in final edited form as: Biochem Biophys Res Commun. 2006 Sep 22;350(2):385–391. doi: 10.1016/j.bbrc.2006.09.069

Fig.2.

Fig.2

TGFβ1 induces ECM gene expression and phosphorylation of Smad3 in adipocytic human HSC. LX-2 cells were cultured in MDI for 72h and TGFβ1 (100 pM) or vehicle were added, then cells were harvested from triplicate wells after 1.5, 3, 6, 12, 24 or 48h. Total protein was extracted and subjected to immuno-blotting for PAI-1 (A, B) and P-Smad3 (E, F). Membranes were stripped and reprobed for β-actin (A) and Smad2/3 (E) to confirm equal protein loading. Total RNA was extracted from parallel triplicate cultures and expression of PAI-1 mRNA (C) and α (1) collagen mRNA (D) were assessed by real time PCR. Results are normalized to the expression of β-actin (for protein studies) or GAPDH (for RNA studies) in the same sample. Reported data are the mean ± SEM of three independent experiments. *p<0.05 versus control.