fig5

Figure 5. Ins-induced collagen synthesis by HSCs is not mediated by the PI3 Kinase signaling pathway. HSCs were isolated from transgenic mice livers and plated as described previously. Cells were grown in high-glucose medium with or without Ins (5 µmol) and PI3 kinase inhibitor wortmannin (W) at a dose of 10 µmol/l. Cells were then harvested for protein extraction and CAT assay on day 7. (A) shows Western blot analysis of AKT phosphorylation by Ins in the absence but not in the presence of wortmannin. Gels were scanned and the density of the test band was divided by its corresponding loading Ctrl, represented as a fold change relative to Ctrl in the accompanying graph; (B) represents the CAT assay on day 7, showing no effect of wortmannin on Ins stimulation of COL1 promoter activity. HSCs: Hepatic stellate cells; CAT: chloramphenicol acetyltransferase; AKT: protein kinase B; Ctrl: control; Ins: insulin; COL1: type 1 collagen.