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. 2020 Jun 12;10:949. doi: 10.3389/fonc.2020.00949

Figure 3.

Figure 3

PdPT induces apoptosis and ferroptosis in non-small cell lung cancer cells. (A) A549 and NCI-H11299 cells were treated with PdPT (5 μM) in the absence or presence of indicated cell death inhibitors for 24 h. BAF, bafilomycin A1 (100 nM); E64d (10 μM); necrostatin-1 (50 μM); zVAD, z-VAD-FMK (50 μM); DFO, deferoxamine (100 μM); Cell viabilities were analyzed by MTS. Mean ± SD (n = 3). *P < 0.05, vs. control. (B) A549 cells were treated with PdPT (5 μM), b-AP15 (1 μM) and RSL3 (10 μM) in the presence or absence of z-VAD-FMK (50 μM), ferrostatin-1 (2.5 μM), or deferoxamine (100 μM) for 24 h. The levels of Annexin V- or PI-positive cells were observed using a fluorescence microscope. (C,D) A549 and NCI-H1299 cells were treated with PdPT (5 μM) in the presence or absence of z-VAD-FMK (50 μM) or DFO (100 μM) for 24 h. The levels of Annexin V- or PI-positive cells were assayed by flow cytometry. Mean ± SD (n = 3). *P < 0.05, vs. control. (E) Western blot analysis of cleaved-PARP and cleaved-caspases in A549, NCI-H1299 and A549/DDP cells following treatment with PdPT for 24 h. (F) Western blot analysis of indicated proteins in A549, NCI-H1299, and A549/DDP cells following treatment with PdPT for indicated times.