Figure 5

Zac1 inhibits cell division and rod fate specification. (a-f) P0 retinae electroporated with pCIG2 control (a-c,e) or pCIG2-Zac1 (d,f) cultured 1DIV. GFP⁺ electroporated cells (green (a-f)) labeled with anti-BrdU (red (c,d)) and anti-ac-3 (red (e,f)). Blue in (b) is DAPI counterstain. (g) Percentage of GFP⁺ cells that incorporated BrdU after electroporation of pCIG2 (black bar; E15.5→1DIV: 88 BrdU/GFP double⁺/542 GFP⁺; P0→1DIV: 124 BrdU/GFP double⁺/1,784 GFP⁺) and pCIG2-Zac1 (white bar; E15.5→1DIV: 24 BrdU/GFP double⁺/290 GFP⁺; P0→1DIV: 14 BrdU/GFP double⁺/816 GFP⁺). (h) Percentage of GFP⁺ cells that expressed ac-3 in P0→1DIV retinae electroporated with pCIG2 (black bar; 157 ac-3/GFP double⁺/5,402 GFP⁺) and pCIG2-Zac1 (white bar; 97 ac-3/GFP double⁺/1,808 GFP⁺). (i-r) P0→8DIV retinae electroporated with pCIG2 (i-k,m,o,q) or pCIG2-Zac1 (l,n,p,r). GFP⁺ electorporated cells (green (i-r)) co-labeled with Pax6 (red; amacrine cells (j-l)), rhodopsin (red; rods (m,n)), Chx10 (red; bipolar (o,p)) and p27^Kip1 (red; Müller glia (q,r)). (s) Percentage of GFP⁺ cells expressing cell-type specific markers post-electroporation of pCIG2 (black bar; 290 Pax6/GFP double⁺/3,939 GFP⁺; 81 syntaxin/GFP double⁺/552 GFP⁺; 955 rhodopsin/GFP double⁺/1,751 GFP⁺; 384 Nr2e3/GFP double⁺/1,261 GFP⁺; 279 Chx10/GFP double⁺/3,146 GFP⁺; 520 p27/GFP double⁺/3,846 GFP⁺) or pCIG2-Zac1 (white bar; 140 Pax6/GFP double⁺/1,284 GFP⁺; 83 syntaxin/GFP double⁺/376 GFP⁺; 56 rhodopsin/GFP double⁺/356 GFP⁺; 131 Nr2e3/GFP double⁺/816 GFP⁺; 263 Chx10/GFP double⁺/1,455 GFP⁺; 541 p27/GFP double⁺/1,888 GFP⁺). Arrowheads mark double⁺ cells. le, lens; Rho, Rhodopsin; Syn, Syntaxin.