Fig. 6: TEAD and coactivator YAP directly bind the SPAG5 promoter.

A Schematic representation of the SPAG5 promoter with the putative TEAD-binding sites (LASAGNA Search 2.0). The promoter regions 657–884 and 2909–3084 were amplified and cloned into pGL3 vector. B, C, E, F Chip analysis of the TEAD and YAP binding on SPAG5 promoter in MDA-MB-231 cell line after TEAD interference detected by qRT-PCR analysis. D–G Transcriptional active chromatin on SPAG5 promoter evidenced by anti-H4-Acetylate antibody. Data are shown as the mean of three independent replicates with the a relative P value. H–J Luciferase assay pGL3 vector carrying SPAG5 657–884 and SPAG5 2909–3084 promoter regions was transfected in MDA-MB-231 cell line after YAP and/or TEAD interference. H MDA-MB-231 cell lines were treated with 0.05–0.1 µM of Dasatinib (I) or with 2 µM of Verteporfin (J) and transfected with pGL3 vector carrying SPAG5 657–884 and SPAG5 2909–3084 promoter regions. Normalized luciferase-activity values and P value from three independent experiments are shown. In the lower panel, western blot shows SPAG5 expression after 0.05–0.1 µM of Dasatinib (I) or with 2 µM of Verteporfin (J) treatments. (ns nonsignificant; *P value <0.05; **P value <0.001).