(A) The number of SCs and total cells were counted at 48 hr post LY+neo and were significantly decreased in lin28a mutant compared with sibling. (B–E) The ET4;ET20 larvae were incorporated with EdU post neo or LY+neo treatment. Three populations labeled with ET4+EdU+ (HC+), ET20+EdU+ (MC+) or ET4-ET20-EdU+ (SC+) were counted and recorded with location information. The proliferative SCs post LY+neo were significantly decreased in lin28a mutant compared with sibling (B and C), while not changed in neo-induced regeneration (E). (D) EdU plots show the positions of EdU+ nuclei of 18 neuromasts superimposed on the same plane, and rose diagrams document the angular positions of SC+. The results show that the proliferative SCs are evenly distributed in each quadrant with no polarization post LY+neo. (F) LY+neo-induced Edu incorporation was significantly reduced in yap mutant. (G) The hs:lin28a larvae were heat-shocked and pre-treated with 10 μM verteporfin before adding neomycin. Samples were collected at 14 hr post LY+neo for EdU+ cell counting. Proliferation (SC+) is significantly decreased post LY+neo in verteporfin, which could be rescued by overexpression of lin28a with hs:lin28a. (H, I) The numbers of regenerated sox2+ progenitors (sox2) and proliferative progenitors (sox2EdU) post LY+neo were both reced in lin28a mutant. Scale bar equals 10 μm.
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