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. 2019 Jun 19;9:484. doi: 10.3389/fonc.2019.00484

Figure 3.

Figure 3

Curcumin-mediated activation of caspase cascade in Pre-ALL cells. Curcumin-induced activation of caspase-9, caspase-3, and cleavage of PARP in Pre-ALL cells. (A) RS4;11 and SupB15 cells were treated with and without 10, 20, and 40 μM of Curcumin for 24 h. Cells were lysed and 50 μg of proteins were separated on SDS-PAGE, transferred to PVDF membrane, and immunoblotted with antibodies against caspase-9, caspase-3, cleaved caspase-3, PARP, and GAPDH. (B) Effect of z-VAD-FMK on curcumin-induced apoptosis and activation of caspase-3. RS4;11, and SupB15 were pre-treated with 20 μM z-VAD-FMK for 1 h and subsequently treated with 20 μM Curcumin for 24 h. Apoptosis was measured with fluorescein-conjugated annexin-V and propidium iodide (PI) and analyzed by flow cytometry. (C) Pre-treatment of z-VAD-FMK prevented curcumin activated caspases. RS4;11 and SupB15 cells were treated as described above and lysed. 50 μg protein lysates were separated by SDS-PAGE, transferred to PVDF membrane, and immunoblotted with antibodies against caspase 9, pro-caspase-3, PARP, and GAPDH. (D) Effect of z-VAD-FMK on CURCUMIN-induced DNA damage. RS4;11 and SupB15 cells were treated as described above and induction of DNA damage was visualized using Comet assay.