Fig. 1. Ethanol exposure induces apoptosis in astrocytes. Astrocytes were exposed to 100 and 200 mM ethanol concentrations for 3, 6, 14 and 24 hours. (A) The sub-G0/G1 astrocyte population, quantified by flow cytometry, increases after ethanol exposure. Representative histograms are shown for control and 200 mM ethanol-treated samples (for 24 hours). (B) Inhibition of the sub-G0/G1 population with the pan-caspase inhibitor z-VAD-fmk (100 µM), at two different ethanol concentrations (100 and 200 mM). (C) Ethanol increases the caspase-3 activity. (D) Preincubation with z-VAD-fmk (50 and 100 µM) abolishes ethanol-induced caspase-3 activation (300 mM for 24 hours). (E) Quantification of early apoptosis using annexin-V together with 7-AAD and detected by flow cytometry. Values are mean ±s.d. of five different experiments. *P<0.05 and **P<0.001 versus control values in one-way ANOVA.