Figure 6.

Cell damage/death of explant human melanoma cultures with lower free zinc stores (M5), average free zinc stores (M10) and higher free zinc stores (M9) exposed to 0.5 μM zinc pyrithione (ZnPyr) during 72 h. Cells (with normal or downregulated expression of Atg5) were treated with zinc pyrithione alone or with pancaspase inhibitor z-devd-fmk, cytochrome c mitochondrial release blocking cyclosporine A, necroptosis inhibitor necrostatin 1 (Nec-1) or autophagy inhibiting 3-methyladenine (3-MA) and their effect on cell damage/death was evaluated by morphometric analysis of cell microgragraphs obtained from time-lapse microscopy (A,B,E,F) or fluorimetric evaluation of active caspase-3 and cleaved PARP-1 presence in the cells (C,D). Results represent means ± SD of five experiments. (A) Phase contrast microscopy, 600×, bar 1 µm. (B) * p < 0.05 significantly higher compared to the beginning of treatment, (F) # p < 0.05 significantly lower than zinc pyrithione only treated cells at the same time interval with one-way ANOVA test and Dunnett’s post-test for multiple comparisons.