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. 2013 Dec 13;289(5):2788–2800. doi: 10.1074/jbc.M113.508143

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FIGURE 3. — Overexpression of ARIA enhances apoptosis in rat neonatal cardiomyocytes. A, isolated rat neonatal cardiomyocytes were transfected with GFP. Immunocytochemistry for GFP and α-actinin and Hoechst staining for nuclei were shown. Bar = 20 μm. B, isolated rat neonatal cardiomyocytes were transfected with either GFP or ARIA-FLAG. Cells were then treated with 50 μm hydrogen peroxide for 12 h. GFP or ARIA-FLAG and Hoechst staining for nuclei were detected under fluorescence microscopy. Apoptotic cells were detected as TUNEL-positive cells. Cells transfected with either GFP or ARIA were analyzed for apoptosis as indicated by arrows. Bar = 20 μm. C, the percentage of apoptotic cells was quantified in rat neonatal cardiomyocytes transfected with either GFP or ARIA-FLAG (n = 4). *, p < 0.05 versus control cells transfected with GFP. D, cells were treated with LY294002 to inhibit the PI3K and then stimulated with 50 μm hydrogen peroxide for 12 h. The percentage of apoptotic cells was quantified in rat neonatal cardiomyocytes transfected with either GFP or ARIA-FLAG (n = 4).