Figure 4.

Knockdown of Taz prevents proliferation, EMT, expression of Zeb1, and Tead1 nuclear localization in cultured RPE cells. RPE cells were infected with either a control lentivirus (A) or a Taz shRNA lentivirus (B), and cells were allowed to grow out in culture for 2 weeks. Lentiviruses expressed GFP, and in the control both infected and uninfected cells proliferated, lost pigment, and underwent EMT. However, the Taz shRNA–infected cells failed to proliferate, lost pigment, and underwent EMT. (C–E) RPE cells were infected with Taz or control shRNA lentiviruses on day 2 in culture, and infected GFP-positive cells were enriched by cell sorting. (C) Live GFP+ Taz shRNA cell is shown to retain epithelial morphology and pigment. (D) Taz shRNA–sorted cells were fixed and immunostained for Zeb1 and Tead1. Fixation eliminated GFP fluorescence. (E) Control shRNA sorted cells were fixed and immunostained for Zeb1 and Tead1. Scale bar, 25 μm.