Figure 4. Interaction of Yap1 with Tead2 is critical for it's ability to bypass Kras^G12D- dependence.

(A) qRT-PCR for expression of Tead family of transcription factors (Tead1-4) in iKras⁻ and iKras⁺ relapse tumors. Error bars represent SD of the mean, ***p < 0.001. (B) Tead2 interacts with Yap1 in Yap1 (Flag-tagged) expressing cells (described in 3C). Input (25%) is used as a reference. (C) Sustained expression of wild-type Yap1 but not TEAD binding defective Yap1 mutants (Yap^S94A and Yap1^Δ60-89) can promote anchorage independent growth of iKras cells offdoxy. For each condition, five random fields were counted. Error bars represent SD of the mean, ***p < 0.001. (D) Mutation in Tead binding domain of Yap1 (S94A) dramatically decreases the ability of Yap1 or Yap1^S127A to substitute for oncogenic Kras in vivo. Representative images shown at 6 weeks off-doxy (n=5 per group). (E) Representative wells (top) of the clonogenic growth assay upon knockdown of Tead2 by two independent shRNAs in Yap1 (or Yap1^S127A) expressing cells (described in 3C). Quantification of cell growth is shown below. Error bars represent SD of triplicate wells, ***p < 0.001. (F) Dominant negative Tead2 (Tead2^DN) selectively suppresses proliferation of Yap1 (or Yap1^S127A) expressing cells but not the Kras^G12D expressing iKras cells. Quantification of cell growth is shown below. Error bars represent SD of triplicate wells, ***p < 0.001. (G) Transcriptionally active form of Tead2 (Tead2-VP16) can substitute oncogenic Kras for in vivo tumor growth. Representative images are shown at 6 weeks off-doxy (n=5 per group).

See also Figure S4.