Pioneer-seq cobinding library, 15 nM FoxA1 + Hnf4a, replicate C
Pioneer-seq cobinding library: a pool of reconstituted nucleosomes carrying a FoxA1 binding site (TGTTTACTTTG, JASPAR MA0148.1) and an Hnf4a binding site (GAGTCCAAAGTCCAG, JASPAR MA0114.2) separated by a fixed 5 bp gap, placed at each of 77 dyad-distance positions across three nucleosomal templates (Widom-601, 5S rDNA, and MMTV-A). The library was incubated with recombinant human FoxA1 and Hnf4a co-titrated at matched concentrations and resolved on native PAGE. The unshifted nucleosome band was excised, amplified, and sequenced on Illumina NextSeq 2000 to quantify per-nucleosome relative shift across three biological replicates.