Figure 3. RASSF1 and RUNX2 restrict YAP1-TEAD oncogenic behavior.

A. U2OS cells were transfected with control vector or plasmid expressing FLAG-YAP1 and indicated siRNAs before colonies were allowed to grow for 11 d, fixed and visualized with crystal violet. Whole cell lysates were probed with indicated antibodies. Error bars indicate SEM of n=3. Significance was determined by student t-Test RASSF1A vs RUNX2/RASSF1A p<0.001; Runx2 vs RUNX2/RASSF1A p=0.023; p73 vs RUNX2/p73 p<0.001. B. U2OS cells were transfected with indicated siRNAs, as shown in A, and cell viability was determined using the resazurin assay after 48 hrs. Error bars indicate SEM of at least n=4. Significance was determined by student t-Test RASSF1A vs RUNX2/RASSF1A p=0.008; Runx2 vs RUNX2/RASSF1A p=0.0101; p73 vs RUNX2/p73 p=0.0185. C. U2OS cells were transfected with indicated siRNAs to promote the different YAP transcriptional partner, as shown in A, and cell proliferation was monitored by xcelligence^TM in real time. Error bars indicate SEM of n=3. Arrows indicate increase or decrease in cell growth, significance determined at end point as below p=0.05 in set of dual knockdown compared to controls.