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Generation of the pomgnt2 line and adult-onset phenotypes in ZKOs. A: Mutation schematic showing indels in exons 1 and 2 induced through CRISPR-Cas9 nonhomologous end joining. B: Western blot showing complete loss of α-DG glycosylation labeled via the glyco-specific IIH6C4 antibody in ZKOs at 1 mpf, even when glycoprotein-enriched lysate is in excess as shown through β-DG protein levels. C: Survival analysis showing ZKOs surviving in Mendelian ratios through 5 mpf. Expected survival rates of 25% for WTs and ZKOs and of 50% for Hets is indicated by dotted lines. D: Body length measurements showing comparable body length between ZKOs and their siblings at 1 mpf, but a significant reduction in ZKOs at 3 and 5 mpf (***p < 0.001). E–G: Images of transverse muscle sections stained for α-DG glycosylation (IIH6C4) (E), F-actin filaments (F), and laminin (G) showing fiber separation, disorganization, and possible fibrosis in ZKOs (20X magnification, scale bars: 100 µm).
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