GSM8721794: Mast cells, F30F R1 001; Mus musculus; OTHER
Bone marrow progenitor cells were harvested from the femurs of 6-8-week-old male WT and MCFosB- mice. Following established protocols, the isolated cells were cultured in a controlled environment at 37°C with 5% CO2.Nuclei were isolated from frozen BMMC pellets. Pellets were homogenized with Dounce homogenizers. Chromatin profiling was performed using the CUT&RUN (Cleavage Under Targets and Release Using Nuclease) technique. Nuclei were isolated from cells and incubated with antibodies targeting specific proteins (e.g., FosB or IgG). Protein A-MNase (micrococcal nuclease) was then added to bind to the antibody-protein-DNA complexes, cleaving DNA fragments at binding sites upon activation with calcium. The released DNA fragments were purified, size-selected, and used for library construction.  Library preparation Sequencing libraries were prepared using the NEBNext® Ultra™ II DNA Library Prep Kit for Illumina® (E7645, New England BioLabs) with NEBNext® Multiplex Oligos for Illumina® (Dual Index Primers Set 1) (E7600S, New England BioLabs) based on manufacture instructions. Libraries were submitted for sequencing by GENEWIZ on Illumina HiSeq 4000 or NovaSeq machine with a 2 × 150-bp paired-end read configuration to a minimum depth of 25 million reads   The libraries were size-selected to enrich for fragments appropriate for paired-end sequencing and quality-checked before sequencing