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. 2019 Nov;189(11):2209–2220. doi: 10.1016/j.ajpath.2019.07.019

Figure 5.

Figure 5

Smooth muscle (SM) α-actin polymerization mediates transforming growth factor-β (TGF-β) and ET-1 signaling. A: Rat hepatic stellate cells (HSCs) were exposed to Ad-control or Ad-SM α-actin [25 or 50 multiplicity of infection (MOI)] for 3 days and incubated in 0.5% serum medium for 24 hours. Cells were harvested in actin stabilization buffer, and F/G-actin isoforms (F/G) were detected by immunoblotting with actin isoform specific antibodies. B–D: Quantification of F/G-actin ratios. E and F: HSCs from Acta2+/+ and Acta2−/− mice were cultured in standard medium for 5 days and then incubated in 0.5% serum medium with or without 20 μmol/L CCG-203971 (CCG) or 1 μmol/L latrunculin (Latr) for 24 hours. Cells were stimulated with TGF-β for 1 hour (E) or ET-1 (20 nmol/L) for 30 minutes (F), respectively. Whole-cell lysates were subjected to immunoblotting as indicated. Representative images and quantitative data are shown. n = 3. *P < 0.05, **P < 0.01, and ***P < 0.001 for Acta2+/+ versus Acta2−/−. cyto–γ-actin , cytoplasmic γ-actin.