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Fig. 4 | Journal of Experimental & Clinical Cancer Research

Fig. 4

From: Hippo component YAP promotes focal adhesion and tumour aggressiveness via transcriptionally activating THBS1/FAK signalling in breast cancer

Fig. 4

YAP-TEAD promoted focal adhesion formation in breast cancer cell lines through inducing FAK phosphorylation. (a) Western blot revealed that compared with the YAP-S94A mutant, overexpression of the YAP-S127A mutant could promote FAK Y397 phosphorylation. (b) Verteporfin reversed YAP-S127A-induced FAK phosphorylation in MCF7 cells. MCF7-YAP-S127A was treated with verteporfin at a dose of 10 μM for 24 h before the Western blot assay was performed. DMSO was used as a negative control. (c) Knockdown of endogenous YAP expression inhibited FAK Y397 phosphorylation in MDA-MB-231 cells. (d) Verteporfin inhibited FAK phosphorylation in MDA-MB-231 cells. Cells were treated with verteporfin at a dose of 10 μM for 24 h before Western blot assays were performed. DMSO was used as a negative control. (e, f) Western blot verified the inhibition of FAK Y397 phosphorylation via defactinib in MCF7-YAP-S127A (e) and MDA-MB-231 (f) cells. Cells were exposed to defactinib at a dose of 10 μM for 8 h before Western blot assays were performed. DMSO was used as a negative control. (g, h) Treatment with defactinib (10 μM) for 8 h decreased focal adhesions, both in MCF7-YAP-S127A (g) and MDA-MB-231 (h). DMSO was used as a negative control. Paxilin (green), F-actin (stained with phalloidin, red). Nuclei (blue). Scale bar: 20 μm. (i) Transwell invasion assays showed that exposure to defactinib (10 μM) could significantly reverse YAP-S127A-induced cell invasion in MCF7 cells. DMSO was used as a negative control. The experiment was performed in triplicate. Scale bar: 100 μm. (j) Exposure to defactinib (10 μM) significantly decreased cell invasion ability in MDA-MD-231 cells. DMSO was used as a negative control. The experiment was performed in triplicate. Scale bar: 100 μm. (k) Quantification of the relative invasion ability in (i). ** p < 0.01 by ANOVA test. (l) Quantification of the relative invasion ability in (j). ** p < 0.01 by Student’s t-test

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