Pr72 KO mediated by TALEN induced cardiac phenotypes in zebrafish. (a) The TALEN target site of exon 1 of the zebrafish pr72 gene. The binding sites used in this study (indicated by “Left” and “Right”) are highlighted in cyan; the SacII site in the spacer is highlighted in red. Representative sequencing results revealed different InDels in the TALEN target site. (b) WISH of cmlc2 expression shows the ventricles and atria in zebrafish embryos, and the ventricles and atria were circled by a dotted line while the ventricle was on the left and the atrium was on the right. The ventricles were enlarged, while the atrial wall was thinning and the atrium was even invisible in the homozygous fish compared to the heterozygous and Wt fish. (c, d) Representative histopathologic sections stained with HE at 40×(c) and 1000×(d), Scale bars 50μM. (c) Representative Hollow arrowheads indicated the ventricular chambers. The bony landmarks pointed by solid arrowheads indicated that the slices were from the same region of the fish body. (d) Higher magnification regions in the red box (1000×). The heart tissues Wt, M1 and M2 were at the same section of the heart. Hollow arrowheads pointed the myocardium nucleus. Red arrows indicated the erythrocytes. (e) Quantification of ventricular chamber areas normalized to ventricular areas, and compared to Wt. (f) Quantification of myocardium cell numbers compared to the control. Myocardium amounts were counted in each of 10 randomly chosen fields per fish at a magnification of 1000× by two observers blinded to the identification of the fish from which the images were obtained. Data are expressed as mean ± SEM. **, p < 0.01 vs controls, one-way ANOVA followed by Dunnett’s posttest. The numbers of zebrafish are indicated in the columns.
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