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Fig. 7. : Cardiac phenotype after loss of Rcsd1 and after rescue with murine Rcsd1 constructs. A. Cardiac phenotype after Rcsd1 depletion and after rescue with murine Rcsd1 constructs at stage 43. The three upper rows show embryos from lateral and ventral view and the respective isolated hearts. The three lower rows show analysed ventricular trabeculation by cardiac Troponin T (Tnnt2) staining. Rows top to bottom: ventral view of the embryos, close-up of the respective hearts and section through the hearts cardiac phenotype after loss of Rcsd1 was rescued by co-injection with the full-length murine Rcsd1 construct and Rcsd1δNLS, although ventricular trabeculation was only partially restored to normal physical appearances. Rcsd1δCPI however was not able to restore cardiac anatomy and physiology. a, atrium; oft, outflow tract; t, trabeculae; v, ventricle, NLS: nuclear localization signal. (Lateral view: red arrowhead highlights cardiac edema upon loss of Rscd1. White arrow heads highlight the cardiac region in embryos of different rescue experiments. Scale bars: upper part: lateral view: 1000 μm; ventral view and isolated hearts: 250 μm; lower part Tnnt2: ventral view: 1000 μm; hearts and sections: 100 μm. B. Quantitative presentation of data shown in A. C. Injection of Rcsd1 MO lead to a reduced heart rate (beats per minute, bpm), which is rescued by co-injecting murine full-length Rcsd1 or Rcsd1δNLS mRNA but not by Rcsd1δCPI mRNA. n, number of independent experiments; N, number of analysed embryos; ng, nanogram. Error bars indicate standard error of the means (s.e.m.). n.s., not significant; **, p≤0.01; ****, p≤0.0001; calculated by a non-parametric Mann-Whitney rank sum test. |
