Figure 5: Inhibition of Hippo protects against I/R injury.
(a,b) Purified adenovirus (1 × 109 opu) was administered by direct injection to the LV free wall (two sites, 25 μl per site) of Tg-DN-Lats2 and WT mice. I/R surgery was performed 5 days after injection. Mice were subjected to ischaemia for 45 min followed by 24 h of reperfusion. (a) Gross appearance of LV tissue sections after Alcian blue and triphenyltetrazolium chloride staining demonstrates that injection of ad-sh-FoxO1 abolished the protection observed in Tg-DN-Lats2 mice. Quantitative measurement of myocardial infarct area/AAR (% infarct size/AAR) was performed (*P<0.05 versus WT/sh-con, #P<0.05 versus Tg-DN-Lats2/sh-FoxO1, n=8). Scale bar, 1 mm. (b) LV myocardial sections were subjected to TUNEL staining. The number of TUNEL-positive myocytes was expressed as a percentage of total nuclei detected by DAPI staining (*P<0.05 versus WT/sh-con, #P<0.05 versus Tg-DN-Lats2/sh-FoxO1, n=5–8). (c–e) Purified adenovirus (1 × 109 opu) was administered by direct injection to the LV free wall (two sites, 25 μl per site). I/R surgery was performed 3 days after injection. Mice were subjected to ischaemia for 30 min followed by 24 h of reperfusion. (c) Sham and ischaemic area samples were used for immunoblot analysis for catalase, MnSOD, and GAPDH. FLAG was also used to examine exogenous YAP gene expression. Statistical analyses of densitometric measurements of catalase and MnSOD are shown (*P<0.05 versus LacZ/sham, #P<0.05 versus LacZ/I/R, n=5). (d) Gross appearance of LV tissue sections after Alcian blue and triphenyltetrazolium chloride staining demonstrates that injection of YAP or YAP S127A decreased the infarct area. Quantitative measurement of myocardial infarct area/AAR (% infarct size/AAR) was performed (*P<0.05 versus LacZ, n=8). Scale bar, 1 mm. (e) LV myocardial sections were subjected to TUNEL staining. The number of TUNEL-positive myocytes was expressed as a percentage of total nuclei detected by DAPI staining (*P<0.05 versus LacZ, n=5–6). Data are shown as mean±s.e.m. P values were determined using one-way ANOVA followed by a Newman–Keuls comparison test.
