Figure 3.

The Zeb1 promoter contains conserved Taz-Tead1 consensus binding sites, Taz binds to the Zeb1 promoter in vivo, and infection with a Taz shRNA lentivirus leads to knockdown of Taz and both Taz and Zeb1 mRNAs. (A) Comparison of human, dog, and mouse Zeb1 5′flanking sequences. Arrow: the transcriptional initiation site. Boxed area: consensus Taz-Tead1–binding sites. Additional consensus binding sites are also evident farther upstream in the promoter. (B) ChIP assay showing binding of both Taz and Yap to the Zeb1 promoter in vivo. IgG indicates immunoprecipitation with a control antibody, and H3 indicates immunoprecipitation for histone H3, a positive control that binds to all genes. (C) Real-time PCR showing the relative levels of mRNA in RPE cells after infection with lentiviruses expressing Taz and control shRNA sequences. (D) Western blot analysis showed that infection with the Taz shRNA lentivirus inhibits the level of Taz but not Yap or β-actin (ACTB).