Supplemental Data from E6 Protein Expressed by High-Risk HPV Activates Super-Enhancers of the <i>EGFR</i> and <i>c-MET</i> Oncogenes by Destabilizing the Histone Demethylase KDM5C
crossref(2023)
Supplementary Figures - Supplementary Figures S1-9. Supplementary Figure S1. Mutual co-immunoprecipitation of HPV16 E6 and KDM5C; Supplementary Figure S2. HPV16 E6 attenuates the KDM5C protein level and KDM5C mRNA quantification; Supplementary Figure S3. HPV16 E6 destabilizes KDM5C and alters the global H3K4 methylation; Supplementary Figure S4. KDM5C does not visibly alters HPV 16E6, HPV 16E7 mRNA and protein levels; Supplementary Figure S5. KDM5C knock out increases tumorigenicity in mice xenograft assay; Supplementary Figure S6. H3K27Ac recruitment to chromatin in the parental cells was examined by the genomic analysis of ChIP-seq signals. ; Supplementary Figure S7. Normalized ChIP-seq signal for H3K27Ac across the EGFR gene and c-MET gene. Supplementary Figure S8. Normalized H3K4me1 levels at KDM5C-bound super enhancers in CaSki-vector and CaSki-KDM5C cells. Supplementary Figure S9. HPV 16E6 depletion leads to EGFR and c-MET super-enhancer inhibition. Supplementary Methods - Description of additional experimental methods including Transfection, siRNA and MG132 treatment, Antibodies, Co-immunoprecipitation and ubiquitination assays, Immunofluorescence Microscopy, Maltose Binding Protein (MBP) Pull-down Assay, Chromatin immunoprecipitation (ChIP)-PCR and ChIP-Seq, Cell Invasion Assay, Real Time Cell Proliferation Assay, Wound healing Assay, Trypsin Digestion and Mass spectrometry analysis Supplementary Tables - Supplementary Tables S1-2. Supplementary Table S1. List of qPCR primers used in this study. Table S2. List of DNA oligonucleotides used in this study.