BAZ1B binds to AT-rich DNA through its AT-hook-like motifs. (A) BAZ1B domain that targets PCH. Schematic drawing of the full-length (FL), truncated BAZ1B consisting of bromodomain (BrD) and three AT-hook-like motifs (BrD+AT1-3), and those motifs’ mutant (BrD+AT1-3mut) is shown, with their representative live-cell images on the right. Contrast was adjusted individually to compare the localization patterns. Amino acid sequence alignments of BAZ1B AT-hook-like motifs in different species and the substituted amino acids in AT1-3mut are shown. (B and C) DNA-binding assay. (B) Experimental design. EGFP-tagged BAZ1B BrD+AT1-3 or BrD+AT1-3mut were expressed in HEK293T cells and purified using anti-GFP nanobody magnetic beads. After incubating BAZ1B domain-bound anti-GFP beads with naked DNA or nucleosomes for 30 min in 20 mM Tris-HCl (pH 7.5) buffer, unbound DNA/nucleosomes were collected. After washing in 20 mM Tris-HCl (pH 7.5) buffer containing 50 mM NaCl, bound-DNA/nucleosomes were eluted using 20 mM Tris-HCl (pH 7.5) containing 350 mM NaCl. (C) Detection of DNA in different fractions by agarose-gel electrophoresis. Relative band intensities are shown below the gels. The positions of size standards are indicated on the right. Scale bar, 10 μm. Source data are available for this figure: SourceData F6.