Figure 4: Hey1 and Id2b regulate MG’s regenerative response.
A, Illustration of hey1 gene promoter from human, mouse and zebrafish with Rbpj binding sites indicated. B, Luciferase assays for Notch (NICD) dependent activation of Wt and mutant hey1 promoter activity; Compared to Wt alone, ***P<0.0001. C, F, EdU Click-iT chemistry on retinal sections identifies proliferating cells in Wt fish treated with indicated MOs (C) or Cas9 and indicated gRNAs (F). D, G, I, Quantification of the number of EdU+ cells at 4 dpi; (D)***P=0.0008, (G) ***P<0.0001, (I) ***P<0.0001. E, H, Quantification of the width of the proliferative zone in retinas at 4 dpi; (E) ***P<0.0001, (H) ***P=0.0004. J, PCR analysis of the effects Hey1 expression has on injury-dependent induction of ascl1a and lin28a. Abbreviations: MO, morpholino; C-MO or Con-MO, control-MO; ONL, outer nuclear layer; INL, inner nuclear layer; GCL, ganglion cell layer; Wt, wild type; m, mutant. Scale bars are 100 μm.
