Fig. 3

Mycb and Mych OE stimulate MG proliferation in the injured and uninjured retina. (A) Bar graph indicating increased MG proliferation (EdU detection, red) at 4 dpi in dorsal and ventral needle poke injured retina with or without Mycb or Mych OE (1 h heat shock repeated every 6 h). Experimental timeline is shown in B. (B) Top: Experimental timeline. Bottom: Representative images used to quantify EdU+ cells as shown in A. (C) Bar graph showing the width of the zone of proliferating MG in the needle poke-injured retina with Mycb or Mych OE as described in A. (D) Experimental timeline is shown at the top. Panels 1a-6a show whole uninjured retina sections taken near the optic nerve head. DAPI-stained nuclei are blue and EdU+ cells are red. Wt and transgenic fish ±DAPT were treated with 1 h heat shock every 6 h, and 3 h before being euthanized fish received an IP injection of EdU. White squares are examples of the dorsal and ventral retinal regions magnified in panels 1b-6b and used for the quantification shown in E. (E) Bar graph showing quantification of EdU+ cells in uninjured retinas treated as in D. (F) Bar graph showing the effects Mycb and Mych OE (1 h heat shock every 6 h) have on MG proliferation (EdU+ cells) in DAPT-treated uninjured retinas with and without Ascl1a/Lin28a OE (1 h heat shock every 6 h). Fish received an IP injection of EdU 3 h before sacrifice on day 4. The number of biological replicates is indicated by the dots in each graph. Error bars are s.d. *P<0.05, **P<0.01. The number of biological replicates (n) is indicated by the dots in each graph. dpi, days post-injury; GCL, ganglion cell layer; HS, heat shock; INL, inner nuclear layer; IP, intraperitoneal; ns, not significant; OE, overexpression; ONL, outer nuclear layer. Scale bars: 50 μm (B); 100 μm (D).