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Fig. 2 Key components of the neuropeptide signaling pathway are expressed in motor neurons (A) Bright-field images of in situ hybridization stainings for cpe and tac1 mRNAs, respectively. Scale bars, 100 μm. (B) Schematic representation of cpe and tac1 gene loci illustrating the CRISPR-Cas9 knockout strategy. Asterisks indicate sgRNA target sites. TSS, transcriptional start site; ATG, start codon; TC, termination codon; PTC, premature termination codon. (C) qPCR showing cpe mRNA levels relative to gapdh. (D) qPCR showing tac1 mRNA levels relative to gapdh. (E) Percentage of wild-type, heterozygous, and homozygous genotypes within a population of 4 dpf cpe or tac1 mutant larvae. In (C) and (D), means ± SEM are shown; Student’s t test. Statistical significance given as ∗∗∗p < 0.001.