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Figure 4. Ins signaling in primary HSCs. (A) depicts AKT phosphorylation in response to Ins. Cells were treated with serum-free medium from days 5-7. On the 7th day, HSCs were treated with Ins (5 µmol) and the proteins were extracted at 15, 30, and 60 min of treatment. Western blot analysis of phosphorylated AKT (normalized to total AKT) showed induced AKT phosphorylation by Ins in HSCs. β-actin was used to demonstrate equal protein loading. Gels were scanned and the density of each band was divided by its corresponding loading Ctrl, and represented as fold change relative to time 0 in the accompanying graph; (B) demonstrates the presence of IRS1 in activated primary HSC cultures, detected by double immunofloresence. The last image shows the co-staining of IRS1 with α-SMA. HSCs: Hepatic stellate cells; AKT: protein kinase B; IRS1: insulin receptor substrate 1; α-SMA: alpha-smooth muscle actin; Ins: insulin; Ctrl: control.