Chip-seq analysis of linker histone-like protein, HP1BP3

HP1BP3 has been subbested to be a linker histone, because it exhibits linker histone-like sequence. To investigate the genome-wide distribution pattern of HP1BP3, Chip-Seq analyses have been conducted. In addition, we found that the function of HP1BP3 as a linker histone was regulated by linker histone chaperon NPM1 and TAF-I. Therefore, we also investigated the effects of histone chaperon knockdwon on the distribution pattern of HP1BP3. As a control, the analyses included the experiments with H1.2, a linker histone variant. Growing HeLa cells were transfected witn either control siRNA, NPM1 siRNA, SET or siRNA. The cells were fixed with 0.5% formardehyde 72 h post siRNA transfection and chromatin was digested by extensive sonication. The extracts were subjected to immunoprecipitation with anti-HP1BP3 and anti-H1.2 antibodies. The preciptiated DNA was extracted and used for libruary preparation for Chip-Seq analysis.