Genome-wide histone modification profiling using ChIP-seq of diabetic or non-diabetic murine bone marrow derived macrophages (BMDM) and haematopoietic stem cells (HSC)

Genome-wide histone modification profiling using ChIP-seq of diabetic or non-diabetic murine bone marrow derived macrophages (BMDM) and haematopoietic stem cells (HSC). Chromatin fragments were selected using anti-Histone H3 (tri methyl K4) (ab8580) or anti-histone H3 (acetyl K27) (ab4729). Sequencing libraries were prepared using the NEBNext Ultra II DNA Library Prep Kit. instructions. The final libraries were purified using SPRI Ampure XP beads to remove adaptor dimers and sequenced by the Biomedical Sequencing Facility (BSF) at CeMM using the Illumina HiSeq 3000/4000 platform and the 50-bp single-read configuration. Overall design: Wild-type C57BL/6J (12-14 weeks old) control or diabetic experimental groups. Diabetes was induced through an intra-peritoneal injection of streptozotocin (STZ) at a low dose range (42 – 45 mg / kg / day) over 5 consecutive days. After 6 weeks post-final STZ injection, diabetic mice with non-fasted blood glucose levels >13.9 mmol/L were sacrificed and tissue collected.