ENU-induced mutations in fugu Mstn gene detected by HRM analysis. The exon 3 of the Mstn gene was examined by HRM analysis with each family of fugu genomic DNA. The region contains the C-terminal peptide of the fugu Mstn protein. The family numbers mean the line of the hatched fry generated by in vitro fertilization (see Table 3). Left side of figures, the high-resolution melting curve (A, C, E, G), and right side of the figures, the substitution plots (B, D, F, H). X-axis and Y-axis, temperature (°C) and relative signal difference, respectively. Red lines and black arrows, the mutations screened by the HRM analysis.