Caspase-3 Deletion Promotes Necrosis in Atherosclerotic Plaques of ApoE Knockout Mice

Figure 1

Apoptosis is inhibited in caspase-3 deficient macrophages and vascular smooth muscle cells. BMDM and VSMCs were isolated from Casp3^+/+ApoE^−/− (Casp3^+/+) and Casp3^−/−ApoE^−/− (Casp3^−/−) mice and treated with cycloheximide (CHX) (0–30 μg/mL) and puromycin (PM) (0–30 μg/mL), respectively. Apoptosis was monitored by (a) western blot analysis for cleaved caspase-3 (β-actin was used as loading control), (b) TUNEL staining ( independent experiments with 2 counting regions of 200 cells/region in duplicate; ^### and ^# versus 0 μg/mL; versus Casp3^+/+; factorial ANOVA with genotype and treatment as category factors; Dunnett post hoc) and (c) caspase-3/7 fluorometric activity assay ( independent experiments; ^##, ^#, and NS, not significant, versus 0 μg/mL; versus Casp3^−/−; factorial ANOVA with genotype and treatment as category factors; Dunnett post hoc).

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