Fig. 8.
Bile acid-induced apoptosis and JNK activation in rat hepatocytes is death receptor independent. Rat hepatocytes were transfected with an adenoviral control AD5LacZ and dominant-negative AD5delta-FADD construct and then treated with GCDC (50 μM), deoxycholate (DCA, 100 μM) for 2 h, or Fas ligand (Fas L; 50 ng/ml) for 4 h. Apoptosis was monitored morphologically by examination of Hoechst-stained cells (A) or biochemically by immunoblotting for CLV3 (B). Cont, control. Equal protein loading was verified by immunoblotting for actin. The amount of phosphorylated JNKthr183,tyr185 was determined by immunoblotting (C). D: rat hepatocytes were pretreated with CPT-2-Me-cAMP (20 μM), BIO (10 μM), or with SB216763 (10 μM) and then exposed to GCDC for 2 h. Cytosolic fractions were prepared by subcellular fractionization as described in materials and methods. The amount of cytochrome c (CYTO C) released into cytosol was determined by immunoblotting. *Significantly different than in the presence of GCDC.