Figure 1.

Actin isoform profiling in Acta2^−/− and Acta2^+/+ hepatic stellate cells. Hepatic stellate cells (HSCs) were isolated from normal Acta2 wild-type (+/+) and Acta2-deficient (−/−) mouse livers and allowed to undergo culture-induced activation as indicated. A: Total RNA was isolated and actin isoform mRNA levels were measured by real-time quantitative PCR using isoform-specific primers. B and C: Cells were isolated as in A, and whole-cell lysates were immunoblotted with specific antibodies. Representative images and quantitative data are shown. Coomassie Blue staining was used to verify appropriate loading (Supplemental Figure S1). D: Cells were isolated as in A and cultured on glass coverslips for 7 days, and then the cells were subjected to immunofluorescence staining. Data are expressed as ΔΔC_t. n = 3. *P < 0.05, **P < 0.01, and ***P < 0.001 for Acta2^+/+ versus Acta2^−/−. Scale bars = 40 μm. cyto–γ-actin, cytoplasmic γ-actin.