Fig. 5: ZFL enhances oxaliplatin-mediated apoptosis.

a Caki cells were treated with 10 ng/mL TNF-α plus 2.5 μg/mL CHX, 500 ng/mL Fas ligand (Fas L), 1 μM doxorubicin, 3 μg/mL etoposide, 250 μΜ 5-FU, 30 μΜ cisplatin, 0.1 μΜ gefitinib, and 5 μΜ sorafenib in the presence or absence of 2 μM ZFL for 24 h. b–e Caki cells were treated with 25 μM oxaliplatin in the presence or absence of 2 μM ZFL for 24 h. We analyzed cell morphology using interference light microscopy and nuclei condensation and fragmentation using DAPI staining (c). Cytoplasmic DNA fragments (d) and caspase activity (e) were detected using a kit, as described in the Materials and methods section. f, g Caki cells were treated with 2 μM ZFL plus 25 μM oxaliplatin in the presence or absence of 20 μM z-VAD-fmk (z-VAD) for 24 h. Flow cytometry was used to detect the sub-G1 fraction, and western blotting was used to detect the protein levels of PARP, pro-caspase-3, cleaved-caspase-3, and/or actin. The values in (a, b, d, e, f) represent the mean ± SD of three independent samples; *p < 0.01 compared to the control; ^#p < 0.01 compared to ZFL plus oxaliplatin