Phosphatidylinositol 3-kinase/Akt pathway regulates hepatic stellate cell apoptosis

Figure 1 Transmission electron micrography of cultured HSC. A: Control HSC showing nuclear is intact and the mitochondria is smooth; B: LY 294002 treated HSC: Chromatins condensed, shrunk and aggregated along inside the nuclear membrane. The arrows points at the apoptotic bodies (× 5000).

Figure 2 Immunocytochemistry (× 400). A: In negative control, the primary antibody was omitted; B and E: PI 3-K p85 and p-Akt⁴⁷³ staining in the PDGF-BB group; C and F: PI 3-K p85 and p-Akt473 expression in the PDGF-BB and LY 294002 groups; D and G: PI 3-K p85 and p-Akt⁴⁷³ staining in the LY 294002 group. Rabbit anti-PI 3-K p85α polyclonal antibody (1:50) and rabbit anti-phospho-Ser⁴⁷³ Akt polyclonal antibody (1:100) used as primary antibodies.

Figure 3 Representative Western blot analysis of PI 3-K protein expression in HSC with β-actin as internal control. From left, 1st lane, control HSC; 2nd lane, PDGF stimulated HSC; 3rd lane, PDGF + LY 294002 group and 4th lane, LY 294002 treated HSC.

Figure 4 Representative RT-PCR photography of PI 3-K mRNA transcription from rat HSC, β-actin as internal control. A: From left, 1st lane, control HSC; 2nd lane, PDGF stimulated HSC; 3rd lane, PDGF + LY 294002 group and 4th lane, LY 294002 treated HSC. B: A graphic analysis of the RNase protection assay.

Figure 5 Representative Western blot analysis of p-Akt and total Akt protein expressions in HSC, β-actin as internal control. From left, 1st lane, control HSC; 2nd lane, PDGF stimulated HSC; 3rd lane, PDGF + LY 294002 group and 4th lane, LY 294002 treated HSC.