Figure 4. Chromatin immunoprecipitation (ChIP) analysis of the HNP-1 promoter in murine and human bone marrow (BM) cells.

BM cells from 3 mice (A–B) and two healthy human donors (C–D) were extracted and fixed in formaldehyde. Cell and nuclear membranes were lysed before fragmentation of DNA by sonication. Chromatin was immunoprecipitated using protein A/G magnetic beads and an antibody against C/EBP-ε, C/EBP-α, or negative control rabbit IgG. After washing procedures, immune complexes were eluted and reversed and DNA recovered. DNA was used as a template for quantitative PCR. Primers used were specific for putative C/EBP sites in the (A,C) HNP-1 promoter and (B,D) murine and human promoters of the specific granule protein cathelin-related antimicrobial peptide (CAMP). Levels are depicted as fold enrichment compared to negative control IgG immunoprecipitation. Error bars depict standard deviation.