Fig. 5

BORIS recruits SRCAP, which drives H2A.Z occupancy at its target sites. a Co-IP: BORIS directly interacts with SRCAP in NIH3T3 + BORIS (clone#2) cells. Benzonase-treated nuclear cell extracts were immunoprecipitated (IP) with either mouse IgG, anti-BORIS Abs, or anti-SRCAP Abs. Western blot analysis with indicated antibodies confirms the interaction. b Average plot of SRCAP occupancy (ChIP-seq data) at BORIS-bound sites in NIH3T3 + BORIS (clone#2) cells compared to NIH3T3 + EV cells. The p-value is calculated by Kolmogorov–Smirnov test. c Heatmap displaying BORIS (left panel, blue), SRCAP (middle panel, purple), and H2A.Z (right panel, brown) occupancy at the 4654 BORIS-bound sites in NIH3T3 + BORIS (clone#2) cells compared to NIH3T3 + EV cells. Arrows atop the heatmaps indicate the proposed sequence of events: BORIS binding to chromatin leads to SRCAP recruitment, resulting in the gain of H2A.Z occupancy around BORIS sites. d Heatmap displaying the correlation between H2A.Z occupancy and BORIS binding in both doxycycline-inducible empty vector (EV) and BORIS-expressing cells. The cells were subjected to treatment with or without doxycycline for a specified duration of time, as indicated