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. 2017 Aug 27;8(43):73407–73418. doi: 10.18632/oncotarget.20562

Figure 1. Sulforaphane targets YAP1/∆Np63α to suppress ECS cell phenotype.

Figure 1

A. B. C. ECS cells were grown for 8 d as spheroids and treated with 0 or 20 μM SFN for 48 h before image acquisition. ECS cells were seeded on a matrigel-coated membrane in a Millicell chamber for invasion assay and then treated with 0 or 20 μM SFN for 20 h. ECS cells were plated as high density confluent monolayers for wound closure assay in the presence of 0 or 20 μM SFN. The values are mean ± SEM and the asterisks indicate a significant reduction (n = 3, p < 0.005). D. SFN treatment reduces YAP1, increases YAP1-P and reduces ∆Np63α. Cells were grown as spheroids for 8 d, treated with 0 or 20 μM SFN for 48 h and lysates were collected for immunoblot. E. F. G. SCC-13 cells were electroporated with control- or YAP1-siRNA and plated for spheroid formation, invasion and migration assay. The values are mean ± SEM and the asterisks indicate a significant reduction (n = 3, p < 0.005). H. YAP1-siRNA treatment reduces YAP1 and ∆Np63α level, but does not impact TAZ or TEAD levels. I. J. SCC-13 cells, electroporated with empty vector (EV) or YAP(S127A), were seeded for spheroid growth or invasion assay in the presence of 0 or 10 μM SFN. Spheroid number was monitored at 6 d. The single asterisk indicates a significant reduction in SFN treated as compared to untreated control cultures. The double asterisks indicate a significant increase as compared to the SFN treated group (n = 3, p < 0.01). K. Immunoblot of extracts prepared from 5 d spheroid cultures (panel I).