Fig. 4

Effects of KC-53 on Caspases and regulatory molecules of the apoptotic pathways in leukemia cells. a Cells were treated with 0 or 5 μΜ KC-53 for the indicated time points. TNFRs membrane expression levels were analyzed by immunoblotting. EGFR was used as a loading control and the TNFRs levels were quantified and normalized in comparison to the EGFR levels. b (i) Cells were treated with 0 or 5 μΜ KC-53 for the indicated time points. (ii) Cells were treated with 0 or 5 μΜ KC-53 in the presence or absence of 20 μM z.vad.fmk. Caspase-8 enzymatic activity was determined as described in Methods. Etoposide (Eto) added at 5 μΜ was used as a positive control. c (i) Cells were treated with 0 or 5 μΜ KC-53 for the indicated time points followed by immunoblotting. (ii) Cells were incubated with 0 or 5 μΜ KC-53 for 6 h and cytosolic and nuclear protein extracts were prepared. The numbers on top of each band represent intensity values and are expressed as fold change in comparison to the control. The results in panels a, b (i) and c are representative of three repetitions. The results in b (ii) panels represent the mean ± SEM of two replicates and are representative of three independent experiments. (***p value <0.001)