The zebrafish wrnKO mutants recapitulate the age-related characteristics of WS patients. (A) qPCR analysis of age-related gene expression (p21 and p53) in intestinal samples at 8 dpf (n = 40 for each group). (B) Western blot images of p53 in WT and wrnKO mutant larvae in intestinal samples at 8 dpf (n = 40 for each group). (C) Viability for WT and wrnKO at 15 dpf, 30 dpf, 45 dpf, and 60 dpf. (D) ORO staining (Top) and WISH of cetp (Bottom) at 8 dpf in WT and wrnKO mutants. Numbers indicate the proportion of larvae exhibiting the expression shown. (E) Representative phenotypic images of WT and wrnKO mutants at the specified time points (15 dpf, 30 dpf, and 45 dpf) and quantification of the body length (n = 12 for each group). (F) Single-optical section images showing the expression of γH2AX (red), Dendra2 (green), and DAPI (blue) in the intestine of Tg(fabp2:Dendra2-NTR) between WT and wrnKO mutants at 8 dpf. Quantification of the percent of γH2AX (n = 10 for each group). (G) Single-optical section images showing the expression of H3K9me3 (red), Dendra2 (green), and DAPI (blue) in the intestine of Tg(fabp2:Dendra2-NTR) between WT and wrnKO mutants at 8 dpf. Quantification of the percent of H3K9me3 (n = 9 for each group). [Scale bars, 1 mm (E) and 50 μm (F and G).] Data are presented as mean ± SEM. Significance was determined by the unpaired t test. NS, not significant; **P < 0.01; ****P < 0.0001. ORO, whole-mount Oil Red O; WISH, whole-mount in situ hybridization.
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