Figure 6
Smad3 linker phosphorylation affects its nuclear translocation in hepatic stellate cells. (A–C) Hepatic stellate cells after passage 1 (HSCs-P1) were transfected with empty vector or with an expression plasmid encoding wild-type or mutant Smad3 (S204A, S208A, S213A) and then treated with R-III (0.25 μM) for 20 h. (A) The treated cells were analyzed by immunofluorescence using anti-Smad3 antibody. (B) Nuclear and cytoplasmic fractions were prepared from the treated cells and subjected to western blotting using anti-Smad3 antibody. Full-length blots are presented in Supplementary Fig. S4. (C) HSCs-P1 were transfected with an expression plasmid encoding wild-type or mutant Smad3 and then treated with or without R-III. The α-SMA and collagen type I (col I) expression levels in the treated cells were analyzed by real-time PCR. P-value, paired t-test (n = 3) (compared with untreated HSCs-P1).
