Figure 4

CAY10594 blocks APAP-induced primary hepatocyte death. (A–C) Primary hepatocytes were isolated from mice liver using collagenase. Cells were preincubated with CAY10594 (10 μM) or not in the absence or presence of 10 mM APAP in a time-dependent manner. (A) LDH activity in the supernatant was assessed by the LDH cytotoxicity assay kit, and OD values at 490 nm are presented as a line graph. (B) To determine hepatocyte death, hepatocytes were stained against propidium iodide (PI) and the number of PI positive cells were counted at x200 magnification by fluorescence microscopy. Scale bar, 100μm (B). (C) Phosphorylation of GSK-3β and total GSK-3β from primary hepatocytes was determined by Western blot analysis. All data are represented as the mean ± SEM. The data are representative of two independent experiments. **P < 0.01; by t-test compared with APAP-treated cells given the vehicle (A).