Figure 5.

YAP mediates the winner versus loser hPSC phenotype

(A–C) YAP displays cytoplasmic localization in crowded wild-type cells co-cultured with variants. YAP (orange) and phosphorylated YAP (p-YAP) (red) staining in wild-type cells in separate culture (A), variant-GFP cells in separate culture (B) and upon co-culture (C). Scale bar: 25 μm.

(D–E) Nuclear to cytoplasmic ratio of YAP (D), and the mean intensity of phosphorylated YAP in the cytoplasm (E). Data are represented as the mean ± SEM.

(F) Representative images of YAP staining in separate cultures. Scale bar: 50 μm.

(G) YAP intensity in variant-GFP and wild-type YAP-overexpressing cells relative to the wild-type cells.

(H) YAP and its target genes are upregulated in wild-type YAP-overexpressing cells compared with wild-type hPSCs.

(I) YAP overexpression leads to improved growth of wild-type cells.

(J and K) YAP overexpression confers the winner phenotype in co-cultures with wild-type cells (J) and increased resistance to cell crowding in co-cultures with variant-GFP cells (K).

(L) YAP intensity in v17q cells transfected with either LUC or YAP siRNA relative to v17q control cells.

(M) Knockdown of YAP in v17q cells co-cultured with wild-type-RFP cells partially rescues the loser phenotype in wild-type-RFP hPSCs.

(G–M) Data are the mean of three independent experiments ± SD. (I, J, K, M), 16 fields were acquired per well.

Statistical analysis was performed by one-way ANOVA followed by Kruskal-Wallis multiple comparisons test (E), Student’s t test (G and L), one-way ANOVA followed by Fisher’s LSD test (H), or two-way ANOVA followed by Holm-Sidak’s multiple comparisons test (I, J, K, M); n.s. nonsignificant; ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001, ^∗∗∗∗p < 0.0001.

See also Figures S5 and S6.