Effect of WNK1 pathway inhibitors on transgene-activated colorectal cancer (CRC) in zebrafish. (A–F) Tg (ifabp:RPIA; myl7:EGFP) adult fish were treated by oral gavage with DMSO (n = 15), WNK463 (n = 15) or Closantel (n = 15) and compared with wildtype fish treated with DMSO (n = 14). QPCR analysis was performed for zebrafish transgene ribose 5-phosphate isomerase A (RPIA), CRC promoting gene cyclin D1 (ccnd1), zebrafish WNK1 orthologue (wnk1a and wnk1b), VEGF receptor-2 (flk1/vegfr2), and angiogenesis/metastasis marker gene matrix metallopeptidase 9 (mmp9). The statistical significance was analyzed by unpaired Student’s t-test. ns: non-significant, p > 0.05; *: 0.01 < p ≤ 0.05; **: 0.001 < p ≤ 0.01; ***: 0.0001 < p ≤ 0.001; ****: p ≤ 0.0001. (G) Representative H&E-stained sections of intestinal tract of RPIA transgenic fish treated with or without inhibitors. Scale bar = 20 μm. (H) Distribution of histopathological findings from tissue sections as shown in panel G. The statistical significance was analyzed by Chi-square test. ****: p ≤ 0.0001.
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