Transgenic rescue of motor coordination deficits in Alpk1PB/PB mice. (A) Diagram of Alpk1 transgene construct. The full-length coding sequence of Alpk1, fused with HA tag, was introduced into the expression vector containing pCX cassette (consisting of a chicken beta-actin promoter and a 5' truncated exon and intron of beta-actin) and a 3' rabbit beta-globin polyadenylation signal. Indicated restriction enzymes ScaI and SfiI were used to excise the transgene from the construct for micro-injection. The transgene-specific PCR primers P8/P9 were used for the transgenic genotyping. (B-E) Analysis of ALPK1 expression in mouse tissues from compound genotypes. Representative western blots showing total ALPK1 proteins in skeletal muscle (B) and brain (D) derived from the indicated genotypes by using anti-ALPK1 antibody. In panel (C, muscle) and panel (E, brain), histogram values represent the ALPK1 densitometric immunoreactive intensity normalized to the intensity of the wild type tissue sample on the same blot.*P < 0.05, ***P < 0.001, n = 3 for each group. (F and G) Behavioral performance of mice with compound genotypes in the dowel test (F) and the rotarod test (G). The motor coordination deficits were completely rescued in the pCX:HAAlpk1;Alpk1PB/PB mice.*P < 0.05, ***P < 0.001, ≥5 per group.