Bulk RNA sequencing SCA1+ fibroblasts from murine skin in the uninjured state and 16 hours, 1.5 days, and 3 days post-wounding

Fibroblasts are emerging as potent modulators of wound-induced inflammation in the skin. We have observed that fibroblasts expressing the surface marker SCA1 are enriched for expression of proinflammatory factors after injury. To characterize how the expression profile of these cells changes as inflammation initiates and resolves following injury, we performed bulk RNA sequencing of isolated SCA1+ fibroblasts from mouse skin in the uninjured state and multiple time points during the post-injury inflammatory phase. Male C57BL/6 mice were used to generate all samples. Unwounded back skin was harvested from three mice for nonwounded (NW) samples 1-3. Full-thickness excisional wounds were made on the back skin of mice using a 4mm biopsy punch and wounds were harvested 16 hours post-wound (hpw), 1.5 days post-wound (dpw), and 3dpw (3 mice per time point). Cells from wound samples and uninjured back skin were isolated with tissue mincing and a liberase digestion solution. Following isolation of the SCA1+ fibroblasts, RNA was extracted from cells and used for sequencing library generation.