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. Author manuscript; available in PMC: 2012 Oct 1.
Published in final edited form as: Dev Dyn. 2011 Sep 8;240(10):2324–2334. doi: 10.1002/dvdy.22734

Figure 5.

Figure 5

Nuclear localization and codon optimization improves function of Gal80. Confocal maximum projections, lateral views, anterior to left, dorsal up, of eyes in Tg(isl2b.3:Gal4)zc65; Tg(UAS:GFP) 72hpf embryos. Scale bar, 50μm. Immunostaining for GFP, green; Topro3 nuclear stain, magenta. (A-A”’) Tg(isl2b.3:Gal4)zc65; Tg(UAS:GFP) transgenic embryos with no Gal80 show GFP expression in all RGCs. Inset and A”’ shows high power magnification of single confocal slice. (B-B”’) Triple transgenic Tg(isl2b.3:Gal4)zc65; Tg(UAS:GFP); Tg(brn3c:Gal80) shows inhibition of Gal4-dependent GFP expression in about 30% of RGCs. Inset and B”’ shows high power magnification of single confocal slice. (C-C”’) Transient injection with construct carrying “improved” Gal80 into Tg(isl2b.3:Gal4)zc65; Tg(UAS:GFP) embryos demonstrates inhibition similar to that of stable lines carrying native Gal80. Improved Gal80 has nuclear localization signal (NLS) and is codon optimized (“opt”).