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. 2020 Aug 10;130(9):4858–4870. doi: 10.1172/JCI134682

Figure 1. Generation of the M213L mouse model and survival of the mutant mice.

Figure 1

(A) Schematic diagram showing the site-directed mutation of ATG, encoding M213, to TTA, encoding an L, at the GJA1 genomic locus using CRISPR technology. HDR: homology directed repair. (B) Representative genotyping results confirmed presence of the M213L mutation in 0, 1, and both copies of the GJA1 gene in WT mice, heterozygous (HT), and homozygous (HM) for M213L, respectively. (C) Quantification of the GJA1 mRNA expression in the hearts of WT, HT, and HM mice. One-way ANOVA, with n = 4 in each group. Data represent mean ± SEM. (D) Representative Western blot showing the expression of GJA1-20k protein in cardiac ventricles of WT, HT, and HM mice (2 to 3 weeks old) after immunoprecipitation. n = 3 experiments. (E) Quantification of GJA1-20k protein expression in cardiac ventricles of 2- to 3-week-old mice among genotypes. Kruskal-Wallis test, followed by Dunn’s multiple-comparisons test, with n = 3 in each group. Data represent mean ± SEM. (F) Kaplan-Meier survival curve of the WT (n = 18, black), HT (n = 36, green), and HM (n = 16, red) mice. Survival curves were compared using log-rank (Mantel-Cox) test.