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

Figure 2

Caspase-3 deficiency in macrophages and vascular smooth muscle cells triggers a switch to necrosis in response to apoptotic stimuli. 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. Necrosis was monitored by (a) morphological analysis (necrotic cells were characterized by cellular oncosis (black arrows) while apoptotic cells showed cellular shrinkage (black arrowheads) and membrane blebbing (open arrowhead)) (scale bar: 25 μm (BMDM) and 50 μm (VSMCs)) and (b) PI labeling ( independent experiments with 2 counting regions of 150 cells/region in duplicate; ^### versus 0 μg/mL; versus Casp3^+/+; factorial ANOVA with genotype and treatment as category factors; Dunnett post hoc).

(a)

(b)