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. 2018 Dec 17;294(7):2302–2317. doi: 10.1074/jbc.RA118.004364

Figure 7.

Figure 7.

SRC-mediated YAP/TAZ activation promotes tumor growth and metastasis. A, A375 cells stably expressing control empty vector or SRCY527F were assayed by Western blotting. B, 5 × 105 cells from A were injected subcutaneously into NOD/Scid mice, and mouse survival was assayed as described under “Experimental procedures.” C, A375 cells expressing a control empty vector, SRCY527F, or SRCY527F and tandem YAP and TAZ shRNAs (shY1/T1) were injected subcutaneously into NOD/Scid mice to assay mouse survival. D, cells from C were also injected into the lateral tail vein of NSG mice, and lung metastases were counted after 21 days. E and F, MA2 cells stably expressing a control shRNA (sh-Control-FF) or an SRC shRNA (sh-SRC313) were assayed by Western blotting (E) or for YAP/TAZ-TEAD transcriptional activity (F). The scatter plot (F) shows mean ± S.D. (error bars), and each dot is an independent experiment in which sh-SRC313 cells were converted to -fold change from control cells (red dotted line). G and H, 5 × 105 cells from E were injected either into the lateral the tail vein (G) or subcutaneously (H) into NOD/Scid mice, and after 5 weeks, lung metastases were counted (G), and primary tumors were isolated and weighed (H). I–L, MA2 or 4T1 cells stably expressing control shRNAs (sh-Control-FF or sh-Control-GFP), SRC shRNAs (sh-SRC313 and sh-mSRC-3), or SRC shRNAs and LATS-insensitive YAP2SA were assayed by Western blotting (I and K) or for YAP/TAZ-TEAD transcriptional activity (J and L). M, 1 × 106 of the MA2 cells from I were injected into the lateral tail veins of NOD/Scid mice, and after 5 weeks, lung metastases were counted. N, 2.5 × 104 of the 4T1 cells from K were injected into the lateral tail veins of BALB/c mice, and after 2 weeks, lung metastatic burden was quantified using qPCR. Scatter plots in D, G, H, M, and N show mean ± S.D., where each dot is a mouse. The plot in F shows mean ± S.D., where each dot is an independent experiment in which the SRC shRNA cells were converted to -fold change from control cells (red dotted line). Statistical significance was tested using Student's t test (D, F, G, and H) and one-way ANOVA with Dunnett's multiple-comparison test (M and N). n.s., p > 0.05; *, p ≤ 0.05; ****, p ≤ 0.0001.