Fig. 6
USP5 deficiency mitigates myocardial reperfusion injury in mice. (A) Representative TTC-Evan’s Blue stained sections of heart from each group. Brick red-stained area represents the area at risk (AAR), whereas the white area indicates the infarcted size (IS). (B) Ratio of AAR/LV, IS/ARR. Data are expressed as mean ± SD (n = 10 for each), and were analyzed using Mann-Whitney U test. *P < 0.05 vs. USP5+/+ +I/R group. (C) Representative cross-sectional images of the left ventricular (LV) showing no-reflow regions indicated by the absence of thioflavin-S fluorescence within the ischemic area that consists of both no-reflow (yellow arrows) and reflow zones (green arrows) following IR. (D) Ratio of no reflow area/LV. Data are expressed as mean ± SD (n = 10 for each), and were analyzed using Mann-Whitney U test. *P < 0.05 vs. USP5+/+ +I/R group. (E) Survival analysis for mice subjected to 45 min ischemia and following 6 h perfusion (*P < 0.05 vs. USP5+/+ +I/R group, Kruskal Wallis test with Dunn post-hoc test for survival analysis). (F-G) Images of echocardiography of mice and quantitative analysis of EF% and FS%. (H-I) Immunoblot analysis and quantification by densitometry of RIPK1, Caspase-8 and c-GSDMD in the heart from USP5+/- mice post-myocardial reperfusion. Data are expressed as mean ± SD (n = 3), were analyzed using Mann-Whitney U test or two-tailed Student t test. *P < 0.05 vs. USP5+/+ +I/R group. (J) LDH activity in serum from USP5+/+, USP5+/-, USP5+/++I/R, USP5+/-+I/R, *P < 0.05 vs. USP5+/+ +I/R group. (K) Heart sections were obtained from USP5+/+ and USP5+/- mice under sham or 45 min ischemia and following 6 h reperfusion. The sections were immunohistologically analyzed by staining with antibody against CD31. (L) Quantitative analysis of inflammatory cells was performed. Data are expressed as mean ± SD (n = 5 for each), were analyzed using Mann-Whitney U test or two-tailed Student t test. *P < 0.05 vs. USP5+/+ +I/R group, Scale bar = 20 μm