Figure 7

(A–M) RT-qPCR analysis of C-type natriuretic peptide system genes during development in Wt zebrafish embryos embryos injected with nppcl/gRNA/Cas9 RNA. Total RNA was extracted from whole TL/Wt zebrafish embryos, which had been reared in either normal aquaria water or exposed to 100 nM CNP, captured at 24, 48, or 72 hpf (n = 3 for each time point) and normalized to 100 ng/µL. (AZD) Target-specific cDNA synthesis was performed followed by multiplex RT-qPCR examining the simultaneous expression of components of the zebrafish natriuretic peptide system (nppc4, nppcl, nppc2, cnp3, nppa, bnp, npra, npr1a, nprb, nprc, corin, furina and furinb). Products were separated using capillary gel electrophoresis and expression quantified using the GenomeLab GeXP analysis system. Data were normalized to the housekeeping gene (elfα) and the mean displayed as fold change over basal expression found in untreated embryos. * p < 0.05, ** p < 0.01, *** p < 0.001 significantly different from control-treated embryos (n = 3 independent experiments).