Analysis of histone H3, H4 acetylation using ChIP assays. (A) Hematopoietic cell lines: MCL Granta (G) and NCEB-1 (N), MM U266 (U), all cyclin D1⁺, and control K562 (K), Manca (M) cell lines. β-Globin 5′ HS2 and glyceraldehyde phosphate dehydrogenase (GAPDH) served as controls. (B) MCL cell line Granta (G) with t(11;14) translocation breakpoint at the P519 region was studied. P519_bkp included a 5′ primer on the translocation breakpoint and a 3′ primer on chromosome 11 so that the PCR detects H3 and H4 acetylation at the P519 region in a translocated chromosome-specific manner. P519_bkp region showed histone H3 and H4 acetylation in Granta cells. Manca (M) was used as negative control. (C) Nonhematopoietic cell lines: H82 (SCLC cell line, cyclin D1 nonexpressing) and MCF7 (breast cancer cell line, cyclin D1 overexpressing). Differential H3 and H4 acetylation was found in the cyclin D1 promoter region, which extended 3 kb upstream and correlated with cyclin D1 gene expression status. All the other regions were acetylated in all hematopoietic cell lines, whereas upstream regions in nonhematopoietic cell lines were hypoacetylated.