Figure 3.

Effects of TF-3 on necroptosis. BEAS-2B cells were treated with CSE (10%)/z-VAD (20 µM) alone or in combination with either TF-3 (20 µM) or necrostatin-1 (20 µM) and were stained with PI and DAPI. (A) For each experiment 100 cells were monitored with a fluorescence microscope. Yellow arrows indicate necroptosis. Scale bars, 20 µm. (B) Quantification of PI positive cells. (C) Cells were stained with PI and Annexin Ⅴ-FITC and analyzed via flow cytometry. (D) Necroptotic rate of cells. Data are presented as the mean ± SEM of three independent experiments indicated by dots. One-way ANOVA was performed followed by Tukey's post hoc test for statistical comparisons among more than two groups. ^**P<0.01, ^****P<0.0001. CSE, cigarette smoke extract; TF-3, theaflavin-3,3'-digallate; Nec-1, necrostatin-1; PI, propidium iodide.