DEV027151 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Subcellular localization of Yap1 in epithelial cell lines. (A-D) In two mouse epithelial cell lines, pam (A,B) and MTD1A (C,D), Yap1 showed strong nuclear signals at low cell density (A,C) and weak nuclear signals at high cell density (B,D). Scale bar in A: 50 µm for A-D. (E,F) Signal intensities of Yap1 proteins in pam (E, n=37) and MTD1A (F, n=45) in the nuclei. Intensities were normalized to the DAPI signal that stains DNA and denoted the value of low cell density as 1. Graphs show averages with standard errors.

• Supplemental Figure S2 -

  Fig. S2. Regulation of Tead1 protein level by Yap1. (A,B) Tead1 protein expression in NIH3T3 cells infected with empty virus (A) or Yap1-S112A virus (B). Virus-infected cells were labeled by EGFP expression. Scale bar in A: 100 µm for A-B′′. (C) Signal intensities of Tead1 proteins in control virus (n=42) and Yap1S112A (n=22) virus-infected cells. Intensities were normalized to the DAPI signal that stains DNA and presented value of control cells as 1. Graph shows averages with standard errors. Overexpression of Yap1-S112A significantly increased level of nuclear Tead1 (P<0.05).

• Supplemental Figure S3 -

  Fig. S3. Differential gene regulation by Tead and Yap1 in NIH3T3 cells, mutant embryos and transgenic mouse. (A,B) qRT-PCR showing no significant alteration of five representative genes identified in NIH3T3 cells in E8.0 Tead1^-/-;Tead2^-/- (A) and Yap1^-/- embryos (B). (C-E) qRT-PCR showing no significant alteration of the genes previously reported as Yap1-regulated genes in transgenic liver and NIH3T3 cells. (C) Previously identified Yap1-induced genes were not induced by Tead2-VP16 or Yap1 in NIH3T3 cells. (D,E) Previously identified Yap1-induced genes were not reduced in E8.0 Tead1^-/-;Tead2^-/- (D) and Yap1^-/- (E) embryos. Graphs show average of two samples with standard errors. (F,G) Comparison of genes regulated by Yap1, Tead2-VP16 and Tead2-EnR. (F) Yap1 and Tead2-EnR affect gene expression in opposing manners. (G) Similarly, Tead2-VP16 and Tead2-EnR affect gene expression in opposing manners.