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. 2023 Jun 15;12(12):e022352. doi: 10.1161/JAHA.122.022352

Figure 2. Apoptosis and necrosis in ex vivo ischemia reperfusion.

Figure 2

A, Uncleaved caspase (Casp) 8 in lysates from control WT (wild‐type) 30‐minute ischemia after reperfusion (IR30) and PI13Kα‐dominant negative hearts (p110α‐DN) IR30 hearts (n=4). B, Uncleaved caspase 3 in lysates from WT IR30 (30‐minute ischemia followed by reperfusion) and p110α‐DN (constituent PI3Kα‐deficient) IR30 hearts (n=4). C, Terminal deoxynucleotidyl transferase‐mediated dUTP nick‐end labeling (TUNEL) staining of the apoptotic nuclei (green) and propidium iodide staining of nuclei (red). D, Bax (bcl‐2‐like protein 4) and Bak (Bcl‐2 homologous antagonist/killer) protein levels in the lysates from nonperfused WT and p110α‐DN hearts (n=6). E, Bax and Bak protein levels in the cytoplasmic (cyto) and mitochondrial (mito) fractions of the lysates from perfused WT IR30 and p110α‐DN IR30 hearts (n=6). F, RIP1 (receptor‐interacting serine/threonine‐protein kinase 1), RIP3 (receptor‐interacting serine/threonine‐protein kinase 3), and pCaMKII (calmodulin‐dependent protein kinase II) protein levels in aerobically perfused WT, aerobically perfused p110α‐DN, 30‐minute ischemia reperfusion WT, and 30‐minute ischemia reperfusion p110α‐DN. Data are presented as mean±SEM; statistical significance is calculated using a 2‐sided t test with Welsch correction in (A, B, D, and E) as well as 1‐way ANOVA in F; *P<0.05 vs WT IR30 (B), *P<0.05 vs WT nonperfused (D), *P<0.05 vs mito WT IR30 (E), *P<0.05 vs aerobic WT (F), # P<0.05 vs WT IR30 (F).