Non specific igg

SiHa cells ectopically expressing ZNF471 were cross-linked with formaldehyde, and chromatin was isolated and sheared by CD-200. Bioruptor® Sonication System (Diagenode, USA) to obtain an average fragment size of 200–500 bp. Sheared chromatin was precipitated with anti-ZNF471 antibody (HPA066695; Sigma-Aldrich, USA). Equal concentration of non-specific IgG (Jackson Laboratory, USA) was used as control. The immunoprecipitated protein-DNA complex was captured using protein A/G immunomagnetic beads (Sino Biological Inc, China), and DNA was isolated by standard phenol chloroform and ethanol precipitation method. PCR was performed to identify the enriched genes (Supplementary Table 4).

Rat anti-mouse ICAM-1 (anti-ICAM) antibody YN1, was produced from respective hybridoma from the American Type Culture Collection (Manassas, VA, USA). Non-specific IgG was from Jackson Immunoresearch (Pike West Grove, PA, USA). 5′-modified DNA oligonucleotide (72-mer) was from Oligo Factory (Holliston, MA, USA). Pierce bond-breaker TCEP solution, LC-SMCC crosslinker, 7 k MWCO Zeba spin columns, thiophilic adsorption resin, heterobifunctional Pierce crosslinking kit, bovine serum albumin (BSA), and trichloroacetic acid (TCA) were from Fisher Scientific (Kerrville, TX, USA). Rabbit anti-mouse PECAM-1 antibody was from Novus Biologicals (Centennial, CO, USA) and FITC-conjugated goat anti-rabbit IgG was from Invitrogen (Carlsbad, CA, USA). Iodogen iodination tubes were from Pierce (Rockford, Illinois) and BioSpin Tris Columns were from BioRad (Hercules, California). Amicon 10 kDa MWCO spin filters were from Millipore Sigma. All other reagents were from Sigma Chemical (St. Louis, MO, USA).