Figure 2—figure supplement 2.

(A) Wnt inhibition with hs:dkk1 has no effect on lin28a induction post neo treatment. (B) Expression levels of lin28a, yap, taz and Hippo pathway genes, such as lats1 and mob1, were detected by western blot at various time points post LY+neo. Results showed that yap was increased at 1 hr post LY+neo. Levels of lats1 and phosphorylated mob1 were not changed at 1 hr, suggesting yap is activated independent of canonical Hippo pathway. (C) Specificity of yap antibody was tested with 32hpf sibling and yap mutants. (D–E) Larvae treated with neo or LY+neo were immonstained with yap antibody. Results showed that nuclear localized yap was increased in LY+neo compared with neo. (F) LY+neo induced yap expression in sibling but not yap mutants. (G) Yap activation post LY+neo was inhibited when atoh1a is inhibited with hs:notch1a, suggesting that yap is activated in atoh1a+ HC precursors. (H) Classic yap target genes cyr61 and ctgfa were upregulated post LY+neo, which is partially blocked by verteporfin. (I) Transcripts of yap and mst2 were highly induced post LY+neo, which was blocked by verteporfin. Scale bar equals 10 μm.