Figure 5

Eye and testicular phenotypes in compound heterozygote Tbc1d20 ^ZFN/bsmice. H&E analysis revealed cataracts in Tbc1d20 ^ZFN/ZFN lenses characterized by the presence of vacuoles (B) when compared to highly organized control lenses (A); scale bars = 50 μm. Tbc1d20 ^ZFN/bs testes appeared smaller in size when compared to controls (C); scale bar = 1 mm. H&E analysis identified disorganized Tbc1d20 ^ZFN/bs seminiferous tubules (E) when compared to highly organized seminiferous tubules in controls (D); scale bars = 50 μm. Immunostaining with TRA54 (green) in control tubules revealed small punctae and crescent-shaped staining consistent with spermatocytes and round spermatids respectively (F) and in Tbc1d20 ^ZFN/bs only TRA54 positive punctate staining was evident (G). PNA positive acrosomes were evident in control tubules (H), whereas in Tbc1d20 ^ZFN/ZFN only PNA positive punctate staining was noted (I); scale bars = 25 μm. DNA was stained with DAPI (blue).