Fig. 11

Effect of SF on cell cycle and apoptotic cell death in HT22 cells. (A) Effect of SF on cell cycle (flow cytometry, stained with PI, n = 3) following treatment with 25, 50 and 75 mM SF for 24, 48 and 72 h. Each value is the mean ± S.D. (B) Effect of SF on cell proliferation (flow cytometry, stained with Click-iT EdU, n = 3) following treatment with 25, 50 and 75 mM SF for 24, 48 and 72 h. Each value is the mean ± S.D. (C) Concentration- and time-dependent effects of SF on apoptosis induction (flow cytometry, labeled with annexin V-FITC and PI) following treatment with 25, 50 and 75 mM SF for 72 h. (D–F) Effects of z-VAD-FMK (D), Nec-1 (E), and Fer-1 (F) on SF-induced cytotoxicity (MTT assay, n = 5) following treatment with 75 mM SF ± z-VAD-FMK or Nec-1 or Fer-1 for 72 h. The cell culture medium was replaced every 24 h.  (G) Protective effect of z-VAD-FMK against SF-induced apoptosis (flow cytometry, labeled with annexin V-FITC and PI, n = 3) following treatment with 75 mM SF ± 5 μM z-VAD-FMK for 72 h. Quantitative values (mean ± S.D.) for different quadrants are shown in Supplementary Fig. S19. Each quantitative value is the mean ± S.D. * or # denotes for P < 0.05 and ** or ## denotes for P < 0.01, respectively.