Fig. 1. Generation and characterization of the R1620L line.
A The R1620L mutation is located in the S4 segment of the DIV transmembrane domain. B Gel electrophoresis image of the digested PCR product (left), confirmed by Sanger sequencing (right). Taq 1 digestion of the PCR product yielded two fragments (610 and 176 bp) from the mutant allele, while the WT allele was identified by the uncut 786 bp PCR product (Fig. 1B). C Homozygous mutants die by postnatal day 22 (P22). Approximately 20% of heterozygous mutants die by P60. D Nav1.6 protein levels were not significantly different between WT littermates, RL/+ mutants, and RL/RL mutants; Kruskal–Wallis test followed by Dunn’s multiple comparisons test. Data are presented as normalized to WT levels with the average WT density = 1. N = 5–7/genotype. Data are presented as mean ± SEM.