Fig. 5.

A competitive interaction between C_Vector and C_YAP cells leads to clonal dominance. (A) Spheroids formed by differentially labeled C_YAP cells (YAP-G or YAP-R) or by differentially labeled C_Vector cells (Vector-G or Vector-R) alone were separately cultured for indicated number of days. Scale bar: 200 μm. (B) Mean±s.e.m. cell numbers in indicated spheroids as shown in A were counted and plotted. n=2 independent experiments. (C) Mean±s.e.m. cell numbers of C_YAP cells in spheroids also containing C_Vector or differentially labeled C_YAP cells as indicated were counted and plotted. n=8 spheroids. (D) Mean±s.e.m. cell numbers of C_Vector cells in spheroids also containing C_YAP or differentially labeled C_Vector cells as indicated were counted and plotted. n=8 spheroids. (E) Cleaved caspase 3 staining was used to detect apoptosis in hybrid spheroids containing indicated differentially labeled C_YAP and C_Vector cells. Representative images from three independent experiments. Scale bar: 50 μm. (F) Mean±s.e.m. numbers of cleaved caspase 3­-positive (CC3⁺) cells from each spheroid in each group as shown in E were quantified. n=10 spheroids. (G) Mean±s.e.m. percentages of CC3⁺ cells overlapped with C_Vector or C_YAP cells (indicated by ZsGreen or mCherry signals) were quantified. For YAP-G:Vector-R, n=9 spheroids; YAP-R:Vector-G, n=8 spheroids; YAP-G:YAP-R and Vector-G:Vector-R, n=6 spheroids. Statistical significance in B–D was calculated by two-way ANOVA and in F,G by Student’s t-test. N.S., no significance; *P<0.05; ***P<0.001; ****P<0.0001.