FIGURE 4

FIGURE 4

The therapeutic modifying effects of fabp7a inhibition were validated in the stable F1 haploinsufficiency mutant. (A) Western blot and quantification analysis of the Fabp7a protein levels in heart lysates of bag3^e2/e2 mutant compared to WT control fish hearts at 6 months. (B) Schematic and chromatographs illustrating the genetic lesion of 8 nucleotides generated by injection of an MMEJ inducing fabp7a sgRNA targeting sequences within the first exon. Dashed lines indicate an 8‐nucleotide‐long deletion. (C) Western blot and quantification analysis of the Fabp7a protein levels in the fabp7a stable heterozygous (fabp7a^e1/+) and homozygous (fabp7a^e1/e1) mutant fish compared to WT controls. n = 3, one‐way ANOVA. (D) Quantification of cardiac function. Ejection fraction (EF) (in %) measured by echocardiography in bag3^e2/e2; fabp7a^e1/+ double mutant fish compared to single mutant and WT control fish at 6 months; n = 6–8, one‐way ANOVA. (E) Confirmative TEM images of bag3^e2/e2; fabp7a^e1/+ double mutant fish hearts compared to bag3^e2/e2 and fabp7a^e1/+ single mutant and WT controls at 6 months. The asterisk indicates Z‐disc aggregation. Arrows point to mitochondrial rounding and swelling. Scale bar: 2 μm. (F) Quantitative RT‐PCR analysis of cardiomyopathy molecular markers in bag3^e2/e2; fabp7a^e1/+ double mutant fish compared to single mutant and WT control fish at 6 months. n = 3 biological replicates, one‐way ANOVA.