Fig. 1: YAP, TEAD, PRC2 and MYC form a nuclear complex onto transcriptionally repressed onco-suppressor genes.

A Chip-Seq tracks of YAP, MYC, TEAD4 and YY1 on the PTEN gene as obtained through the Cistrome Browser DB tool and displayed in the WashU Epigenome browser. MYC, YY1 and TEAD4 tracks were obtained in the A549 cell (https://genome.ucsc.edu/cgi-bin/hgTrackUi?db=hg19&g=wgEncodeHaibTfbs) [47]. YAP track was obtained in the NCI-H2052 cell line [48]. Red lines indicate the regions amplified for ChIp analysis. Chromosomal coordinates are relative to the Genome Reference Consortium Human Build 38 (hg38). B Chromatin immunoprecipitation analysis of the abundance of MYC, YAP and YY1 in H1299 cells depleted for MYC, YAP and YY1, respectively, compared to siGFP as control. Fold enrichment was calculated over IgG and then normalized to the siGFP control that was adjusted to 1. The experiments were performed in triplicate. An intronic region of the EZH2 locus was used as a negative control for MYC and YY1 binding, while CTGF promoter was used as a positive control for YAP binding. Two-tailed t-test analysis was applied to calculate the P values. *p < 0.05; **p < 0.01; ***p < 0.001. p and n values: PTENp1 siMYC vs siGFP p = 0.08, n = 3. PTENp2 siMYC vs siGFP p = 0.004, n = 3. neg ctrl siMYC vs siGFP p = 0.1, n = 2. CTGF siMYC vs siGFP p = 0.37, n = 2. C H1299 protein lysates were immunoprecipitated (IP) with anti-IgG, anti-YAP, anti-pMYC or anti-YY1 antibodies and then subjected to Western blot (WB) with the indicated antibodies (right side). 5–10% of total lysate was used as input control. Confocal analysis of YAP (green) and Lamin A/C (red) signals of H1299 cells seeded at higher density (D) or lower density condition (E). Magnification = ×63. Scale bar = 22 µm (D), 10 µm (E). F Confocal analysis of pMYC (green) and Lamin A/C (red) signals in H1299 cells seeded at high density. G–J Confocal analysis of the indicated proteins in H1299 cells seeded at low-density culture conditions. Magnification = ×63. Scale bar = 17 µm (F) and 10 µm (G–J). For each analysis, DAPI (blue) and merge are shown. K PLA for the analysis of the interaction between proteins indicated in panels. Magnification = ×63. Scale bar is indicated in panels.