Fig. 6.

Role of Akt and FOXO3a in ALK4/5-mediated HK-2 cell death induced by cadmium. a–c Cells were incubated with 0.1% DMSO, 10 μm SB431542, 10 μm SB505124 a, or 10 µm MK2206 a–c for 1 h and then incubated with or without 25 μm CdCl₂ (Cd) for 15 h a or 30 h b, c. Cell lysates were subjected to western blotting using the indicated antibodies a. The viability of cells was determined by trypan blue exclusion assay b. Percentage of propidium iodide (PI) or Annexin-V (AV)-positive cells were determined by Annexin-V and PI staining c. d Cells transfected with control siRNA or FOXO3a siRNA were incubated with 0.1% DMSO or 10 µm SB431542 for 1 h and then incubated with or without 25 μm CdCl₂ (Cd) for 36 h. The viability of cells was determined by trypan blue exclusion assay. Immunoblots shown are representative of at least three independent experiments. Each value of trypan blue exclusion assay is the percentage of trypan blue-positive cells and reflects the mean ± SD of at least three experiments with duplicate assays in each experiment. Results of Annexin-V and PI staining are representative of at least three independent experiments. *P < 0.05, **P < 0.01, significant difference between the samples