Fig. 1.

stk3 mutation induced edema, glomerular cyst and pronephric tubule dilation in zebrafish. (A) Schematic diagram of stk3 gene structure and CRISPR/Cas9-induced mutation. The CRISPR site was designed to target the 6th exon. A mutant line with a 5-bp insertion was generated for phenotype analysis. Filled boxes indicate coding regions and open boxes indicate untranslated regions. The underlined sequence is the CRISPR site, the protospacer adjacent motif (PAM) sequence is shown in blue, and the altered sequence is shown in red. (B) Morphology of larvae at 15 dpf. The asterisk indicates the edema. Scale bar: 1 mm. (C) Kaplan–Meier plots for the onset of edema in stk3^−/− larvae and controls [wild-type (WT) and heterozygous siblings]. All the stk3^−/− larvae exhibited edema during 12-18 dpf, with median onset time at 14.5 dpf. Statistical analysis was performed using log-rank (Mantel–Cox) test (P<0.001). (D) Kaplan–Meier plots for survival of stk3^−/− larvae and controls (WT and heterozygous siblings). No stk3^−/− larvae could survive beyond 20 dpf, with median survival time at 15 dpf. Statistical analysis was performed using log-rank (Mantel–Cox) test (P<0.001). (E) Schematic diagram of larval cross section. (F) H&E staining of paraffin-embedded sections of stk3^−/− larvae and WT siblings at 15 dpf. The boxed areas are shown enlarged below. stk3^−/− larvae exhibited glomerular cysts with enlarged Bowman's space (asterisks) and pronephric tubule (arrowheads) dilation. Scale bars: 100 μm (top) and 50 μm (bottom).