Figure 3
CASPASE-9, CASPASE-3 activation, and PARP cleavage upon 1% O2 hypoxia induction. (a) Cleavage and activation of initiator CASPASE-9, effector CASPASE-3, PARP proteolysis (CASPASE-3 substrate), and HIF-1α stabilization were analyzed by western blot in H9 hESCs and FN2.1 hiPSCs at 4-, 8- and 24-h post 1% O2 incubation. ACTIN was used as loading control. Representative blots of three independent experiments are shown (original images are presented in Supplementary Fig. S10 and S11). (b) CASPASE-3 activity was determined upon 8-, 16- and 24-h of 1% O2 treatment by measuring the proteolysis of CASPASE-3 substrate Z-DEVD-R110. Specific activity was calculated as fluorescence/number of cells and expressed as fold induction against normoxia. Mean + SEM fold induction relative to normoxia (arbitrarily set as 1) of three independent experiments is shown. Statistical analysis was done by Student’s t-test, (*) p < 0.05 and (**) p < 0.01 versus normoxia.
