The increasing extents of H2O2 levels were significantly higher in the injury Δ113p53M/M mutant hearts.(a–d) After surgery, Tg(Δ113p53:GFP) animals were treated with either DMSO (a, a′, c, c′) or DPI (b, b′, d, d′) daily at 3–7 or 7–14 dpa. The treated animals were sampled at 7 and 14 dpa and subjected to cryosection. Cryosections of hearts were immunostained by anti-GFP (in green) and anti-MHC (MF20) (in red) antibodies. The nuclei were stained with DAPI (in blue). Scale bar, 50 μm. e Average size of GFP+ cardiomyocytes on heart sections of Tg(Δ113p53:GFP) treated with DMSO or DPI at 7 and 14 dpa, was presented as the percentage of the ventricular area at the resection site. Data are means of three sections/heart. Each dot represents the average size of GFP+ cardiomyocytes in an individual heart. Scale bar, 50 μm. f–l Ex vivo HyPer heart images of either Δ113p53+/+ (f, h, j) or Δ113p53M/M mutant hearts (g, i, k) at sham, 10.5 and 21 dpa. Spatially resolved H2O2 image, indexed by the ratio between the F488 and F405 images of HyPer (below), is presented in pseudocolor. Ratiometric HyPer signals (F488/F405) averaged over the regenerative zone of injured heart at 3.5, 7, 10.5, 14, 17.5 and 21 dpa were presented as the difference to the average F488/F405 ratio at the apex of respective sham hearts (l). Each dot represents the ratiometric HyPer signal in an individual heart. Statistical analyses were performed on data from Δ113p53+/+ and Δ113p53M/M mutant hearts at the same time point. The experiments were repeated independently for at least three times with similar results. Statistical analysis was performed on relevant data using Student’s two-tailed t test in GraphPad Prism 5. The p values were represented by n.s. and asterisks. n.s., p > 0.05. *p < 0.05. **p < 0.01. ***p < 0.001.
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