Analysis of the expression profiles of back skin and excisional wound-derived fibroblasts FACS-sorted from mice overexpressing activin A.

Overexpression of activin A by keratinocytes accelerates excisional wound healing in mice. Activin-promoted wound healing is mediated via the stroma, specifically by the wound immune cells and fibroblasts. To determine if activin A alters the gene expression profile of wound fibroblasts, we performed RNA-sequencing of fibroblasts FACS-sorted from excisional skin wounds of activin overexpressing mice. We found that activin induces a pro-fibrotic gene expression profile of these fibroblasts, leading to the upregulation of genes involved in collagen biosynthesis and remodeling. CD45-negative/CD140a-positive live cells (fibroblasts) were FACS-sorted from back skin (BS) and excisional skin wounds (5-days post-injury, 5dw) of activin overexpressing mice (K14-Act) and their wild-type (WT) littermates. The experiment was performed in triplicates; for each replicate, material from 3 mice of corresponding genotype and tissue was pooled. The expression profiles of the sorted fibroblasts were analyzed by RNA-sequencing.