GSM8784203: Macrophages, Glscko2, Negative Control; Mus musculus; OTHER
Macrophages were isolated from wound bed skin tissue using the anti-F4/80 MicroBeads UltraPure, mouse (Miltenyi Biotec, Catalog #130-110-443) according to the manufacturer's protocol. Briefly, 6 wounds from 3 mice were pooled for each sample and a total of 6 samples were used for each timepoint. Wound bed tissue was excised, minced, and digested with Collagenase IA and Collagenase II to obtain a single-cell suspension, as previously described. After filtration through a 70-µm cell strainer to remove debris, cells were washed and resuspended in MACS buffer (PBS containing 0.5% BSA and 2 mM EDTA). The single-cell suspension was incubated with anti-F4/80 MicroBeads for 15 minutes at 4°C. Labeled cells were then passed through an LS column (Miltenyi Biotec, Catalog #130-042-401) placed in a magnetic field using the QuadroMACS™ Separator (Miltenyi Biotec, Catalog #130-090-976) with MACS® MultiStand (Miltenyi Biotec, Catalog #130-042-303) according to manufacturer directions. CUT&Tag libraries were prepared using the CUTANA™ CUT&Tag Kit (Epicypher, SKU: 14-1102) following the manufacturer's instructions