Figure 4.

NO donors did not affect the inhibition of TGFβ1-activated HSCs by BAY 41-2272. HSCs were preincubated with BAY 41-2272 (10 µM) in the presence or absence of SNAP (0.1 µM) or SNP (0.1 µM) for 30 min before activation by TGFβ1 (5 ng/mL) for another (A–C) 6 h or (D) 24–48 h. (A) mRNA levels of CTGF and GAPDH were determined by quantitative RT-PCR. (B,C) Expressed CTGF and GAPDH proteins were measured by (B) Western blot and (C) immunofluorescent staining using the corresponding antibodies. (D) Cell viability was measured using the WST-1 assay and monitored spectrophotometrically at 450 nm. All data are expressed as mean ± SD (n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001 compared with the TGFβ1 alone.