Figure 1. Generation and characterization of E2F5/p130 double transgenic mice. A: Microinjected constructs. Human E2F5 and mouse p130 cDNAs were linked to the δenhancer αA-crystallin promoter in vector δenαA-minx (DREAM) [30]. B: At E15.5, the bigenic lens shows a normal lens phenotype. C: No BrdU positive fiber cells were present in the center of the double transgenic lens. D-E: In situ hybridization was used to examine expression of E2F5 and p130 transgenes with human E2F5 and mouse p130 riboprobes. Hybridization signals were initially captured as dark-field images, pseudocolored red, and then superimposed on bright-field images of the same tissue sections counterstained by hematoxylin. The E2F5 (D) and p130 (E) transgenes were expressed in lens fiber cells. The size of lens in panel E is smaller as this section is more peripheral. F-G: Immunohistochemistry. Using antibodies against E2F5 (F) and p130 (G), a green nuclear staining was detected in the double transgenic lens fiber cells. Scale bars=500 μm.