Fig. 4: MYC facilitates CTCF chromatin binding.

a Multi-omics assays to evaluate the function of MYC on CTCF binding and looping in 22Rv1 cells. b Western blot assay showing the efficiency of MYC overexpression. c The overlap of MYC, CTCF and H3K27ac peaks between control (Ctrl) and MYC overexpression (MYC-OE) cells. d Heatmaps showing the ChIP-Seq signal of MYC, CTCF and H3K27ac at MYC and CTCF peaks. From top to bottom, MYC and CTCF peaks were separated into shared, MYC-only, and CTCF-only groups. e The overlap between MYC and CTCF peaks in MYC-OE cells. f The aggregated CTCF ChIP-Seq signal in Ctrl and MYC-OE cells. CTCF peaks were divided into four groups based on MYC and H3K27ac status. g The aggregated H3K27ac ChIP-Seq signal in Ctrl and MYC-OE cells. H3K27ac peaks were divided into four groups based on MYC and CTCF status. h Co-immunoprecipitation to detect the protein-protein interaction between CTCF and MYC in Ctrl and MYC-OE cells. i Western blot and corresponding Coomassie blue staining of GST pull-down assay. 22Rv1 cell lysates were subjected to pulldowns with immobilized GST only or recombinant GST-CTCF protein. Bound protein was probed with anti-CTCF and anti-MAX antibodies by Western blot. The red arrow indicates the GST-MYC band. j 22Rv1 cells expressing GFP-MYC and Flag-CTCF were used in co-IP assay. The input and IPed proteins were analyzed by Western blot with anti-GFP and anti-Flag antibodies. k Proximity Ligation Assay to detect the in-situ interaction between MYC and CTCF proteins. Nuclei were stained with DAPI. Scale bar, 10 μm. For b, h, i, j, and k, these experiments were repeated independently three times with similar results. Source data are provided as a Source Data file.