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. Author manuscript; available in PMC: 2018 Oct 14.
Published in final edited form as: Nat Cell Biol. 2015 Aug 10;17(9):1218–1227. doi: 10.1038/ncb3216

Figure 3. Control of cell proliferation by YAP/TAZ/TEAD.

Figure 3

(a) Growth curve of MDA-MB-231 cells transfected with control siRNA (siCO) or two different combinations of siRNAs targeting YAP and TAZ (siYT). Data are mean+SD of n=8 biological replicates. Individual depletion of YAP or TAZ has no effect on cell growth (Supplementary Fig. 3a).

(b) Percentage of MDA-MB-231 cells in G1, S and G2/M phases of cell cycle, as determined by flow-cytometric analysis of DNA content. Cells were transfected with control (siCO) or YAP/TAZ siRNAs (siYT) 48hr before fixation. Data are mean+SD of n=3 biological replicates.

(c) Sustained expression of TAZ, but not of TEAD-binding deficient TAZS51A, rescues cell proliferation in YAP/TAZ-depleted cells. Empty-vector-, wild-type TAZ- (wt) or TAZS51A-transduced MDA-MB-231 cells were transfected with control (siCO) or YAP/TAZ (siYT) siRNAs, as indicated. Proliferation was evaluated as in (f). Data are mean+SD of n=8 biological replicates.

(d) MDA-MB-231 cells were transduced with lentiviral vectors encoding rtTA and doxycycline-inducible MYC (MDA TetON MYC) and transfected with control or YAP/TAZ siRNAs. Where indicated, MYC expression was induced with 0.1 μg/ml doxycycline at the time of transfection. Cell growth was evaluated as in (f). Data are mean+SD of n=8 biological replicates. A control experiment with doxycycline-inducible EGFP is shown in Supplementary Figure 3h.

(e) Average gene expression values of validated YAP/TAZ/TEAD direct target genes (listed in Fig. 2b) in invasive breast cancer samples, classified according to their histological grade. Individual data points and the mean value (black line) of each group are shown.

(f) Kaplan-Meier graph representing the probability of cumulative metastasis-free survival in breast cancer patients stratified according to the expression of validated YAP/TAZ/TEAD direct target gene signature. High expression of the signature is associated with shorter metastasis-free survival (log-rank p-value < 0.0001).

See Methods for reproducibility of experiments.