Figure 3.

Cells were treated with or without CB-PIC (0 or 40 μg/mL) under normoxic or hypoxic conditions for 6 hr. (a) Cell lysates were prepared and subjected to Western blotting to determine the expression of AMPKα, pAMPKα, PARP, BCl-2, pERK, ERK, and β-actin. Band density of pAMPKα, cleaved PARP, BCl-2, and pERK were quantified using Gel-pro analyzer (Media Cybernetics, Bethesda, MD, USA). Values are means ± SD, n = 3. *P < 0.05 and ***P < 0.001 compared with normoxia and hypoxia groups. (b) Cells (1 × 10⁶ cells) treated with CB-PIC for 6 hours were measured enzyme activity of the caspase-3 class of protease in apoptotic cells by using Caspase-3 Colorimetric Assay Kit. (c) Cells were transiently transfected with AMPKα siRNA or control siRNA in the presence or absence of CB-PIC (40 μg/mL), and PD 98059 was also treated to SW 620 in the presence or absence of CB-PIC (40 μg/mL) for 6 hours in hypoxia. Western blotting was performed to determine the expression of PARP, AMPKα, pAMPKα, pERK, ERK, and β-actin. (d) TUNEL staining was performed and visualized under fluorescence microscopy (×200).