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Figure 1. HEMA-induced apoptosis and caspases activation. (A) Dose-dependent induction of apoptosis. Cells were treated with various concentrations of HEMA for 24 hrs, and apoptotic and necrotic cells were stained with annexin V-FITC or PI, respectively. Cells were then detected and quantified by flow cytometry as a percentage of the entire population (see MATERIALS & METHODS). Results represent the means ± SEM of 4 independent experiments in duplicate (n = 4). * Significantly different from the untreated control group (one-way ANOVA followed by Bonferroni post hoc test, p < 0.05). (B) Dose-dependent caspase-3 activation. Cytosolic extracts from cells treated with 0–10 mM HEMA for 24 hrs were analyzed by Western blotting with anti-caspase-3 antibodies. (C) Time-dependence of caspase-3 activation. The cells were treated with 10 mM HEMA for the indicated periods of time, and cytosolic extracts were analyzed by Western blotting with anti-caspase-3 antibodies. (D) Caspase-8 and -9 activation. Cytosolic extracts from cells treated with 10 mM HEMA for 24 hrs were analyzed by Western blotting with anti-caspase-8 and -9 antibodies. The amounts of cell lysate were normalized with the use of polyclonal anti-actin or tubulin antibodies. Each Western blot is representative of at least 2 independent experiments.
