FIGURE 2.

Yun is required for tumorigenesis. (A) Expression of Raf ^gof in esg ⁺ cells causes tumours (green, by esg > GFP) at 29°C for 7 days (white arrowheads). (B) The number of tumour cells in esg ^ts  s Raf ^gof intestines (green, by esg > GFP) is dramatically decreased by co‐expression of yun ^shRNA at 29°C for 7 days (white arrowheads). (C) Quantification of the relative number of ISCs in esg ^ts  > Raf ^gof intestines and esg ^ts  > Raf ^gof , yun ^shRNA intestines. Mean ± SD is shown. n = 15. ****p < 0.0001. Please note that esg ^ts  > Raf ^gof intestines are highly deformed which prevent the accurate quantification of esg ⁺ cells in esg ^ts  > Raf ^gof intestines. (D) pH 3 (red) in esg ^ts  > Raf ^gof intestines at 29°C for 7 days (white arrowheads). (E) pH 3 (red) in esg ^ts  > Raf ^gof , yun ^shRNA intestines at 29°C for 7 days (white arrowheads). (F) Quantification of pH 3 number per gut in esg ^ts  > Raf ^gof intestines and esg ^ts  > Raf ^gof , yun ^shRNA intestines. Mean ± SD is shown. n = 15. ****p < 0.0001. (G) Expression of Raf ^gof in ISCs increases ISC number at 29°C for 7 days (white arrowheads). (H) The number of ISCs in Dl ^ts  > Raf ^gof intestines is significantly decreased by co‐expression of yun ^shRNA at 29°C for 7 days (white arrowheads). (I) Quantification of the relative number of ISCs in Dl ^ts  > Raf ^gof intestines and Dl ^ts  > Raf ^gof , yun ^shRNA intestines. Mean ± SD is shown. n = 15. ****p < 0.0001. (J) MARCM clones of Raf ^gof (green) at room temperature for 6 days after clone induction (ACI). (K) The size of Raf ^gof clone is totally suppressed in yun ^Δ9 (ACI 6 days). (L) Quantification of the clone size of indicated genotypes. Mean ± SD is shown. n = 15. ****p < 0.0001. In all panels except graphs, GFP is in green and blue indicates DAPI staining of DNA. Scale bars: 20 μm