Fig. 3

NYT inhibited CSE-induced caspase-dependent apoptosis. The effects of specific inhibitors of apoptosis and necrosis were evaluated using an LDH assay kit. Two hours before exposure to 5% CSE, the cells were treated with either Z-VAD-FMK (A) or necrostatin-1 (B) to inhibit apoptosis and necrosis, respectively. The cytotoxicity of LDH leakage was evaluated 24 h after CSE exposure. ^###p < 0.001 vs. control group; ^**p < 0.01, ^***p < 0.001 vs. vehicle-treatment group; Tukey’s t-test; n = 6. The effect of NYT on the expression levels of cleaved caspase 3 and PARP was evaluated by Western blotting analysis (C-E). Twenty-four hours after CSE exposure, the cells were collected, and western blotting analysis was performed to evaluate the expression levels of cleaved caspase 3 (D) and cleaved PARP (E). The full-length blot images were presented in Supplementary Fig. 1. ^*p < 0.05 vs. vehicle-treatment group; Student’s t-test; n = 3. CSE: cigarette smoke extract; NYT: Ninjinyoeito