Fig. 4
From: PHF6 recruits BPTF to promote HIF-dependent pathway and progression in YAP-high breast cancer
YAP signals govern the high PHF6 expressions in breast cancer cells. A The RT-qPCR analysis of PHF6 mRNAs in MCF-7 cells treated with DMSO and indicated compounds targeting various signals. The levels were normalized and illustrated by heatmap. B Western blotting assay detecting the PHF6 proteins in MCF-7 cells transfected with EV, YAP-S127A and YAP-S94A plasmids. C Western blotting assay detecting the PHF6 proteins in shCtrl and shBPTF MDA-MB-231 cells. D, E Dual luciferase reporter assays showed that PHF6 promoter activity in MCF-7 (D) and MDA-MB-231 (E) cells tranfected with EV, YAP, YAP-S127A and YAP-S94A plasmids. F ChIP-qPCR assay was conducted with a-YAP antibody and control IgG to show YAP binding to PHF6 promoter in parental and YAP-deleted cells. G ChIP-qPCR assay was conducted with a-TEAD4 antibody and control IgG to show TEAD4 binding to PHF6 promoter in parental and YAP-deleted cells. Besides, YAP loss impaired TEAD4 binding ability. H Detection of relative luciferase activity in the indicated YAP-deleted MCF-7 cells transfected with EV-GFP, YAP, TEAD4, YAP-S127A and YAP + TEAD4, respectively. I CCK-8 assays were utilized to determine cell growth rates in the indicated groups, including EV + shCtrl, YAP + shCtrl, and YAP + shPHF6#1. MCF-7 cells (left), MDA-MB-231 cells (right). J Correlation analysis showing the relationships between PHF6 and YAP1 levels in TCGA-Bca cohort. K The RT-qPCR analysis of PHF6 mRNA levels in shCtrl and shYAP MCF-7 cells that were exposed to 20% or 1% O2 for 24 h, individually. L The RT-qPCR analysis of PHF6 mRNA levels in parental and HIF-DKO Bca cells. M Western blotting assays were used to detect PHF6 proteins in Bca cells under 20% or 1% O2 for 24 h, individually. *P < 0.05, **P < 0.01, ***P < 0.001