Fig. 5: Adipocytes are non-fibrogenic in wounds.

mRNA-seq was performed with FACS sorted adipocytes and fibroblasts from day 7 and 21 wounds and adjacent skin of Adipoq^Cre;R26^mTmG and En1^Cre;R26^mTmG mice, respectively. Each cell type at each time point includes three independent biological replicates. a. Pearson correlation analysis of all 18 samples. Colour in each cells represented Pearson correlation coefficients for every pairwise comparison. b GO term enrichment based on DEGs of adipocytes and fibroblasts from day 10 wounds. Filled colour represented number of genes enriched relative to all DEGs. Cryosections of day 7 and day 21 wounds from Adipoq^Cre;R26^mTmG mice were subjected for immunofluorescence staining. c Representative images and quantification of Perilipin (magenta) in GFP positive cells. Data are numbers of GFP⁺Perilipin⁺ cells per high magnification field, n = 6 independent samples, mean ± SD, unpaired two-tailed t-test. d Representative images and quantification of αSMA (magenta) in GFP positive cells. The migratory elongated and rounded adipocytes are negative for αSMA. At day 7 there is widespread αSMA staining in the centre of the wound, whereas only physiological αSMA is found in the hair follicle dermal sheath at day 21. Data are percentage of αSMA+GFP+ cells and αSMA-GFP+ cells in total GFP+ cells, n = 6 independent samples, mean ± SD. e Representative images and quantification of vimentin (magenta) in GFP positive cells. Data are percentage of Vimentin⁺GFP⁺ cells and Vimentin^-GFP⁺ cells in total GFP⁺ cells, n = 6 independent samples, mean ± SD. f Transplantation of FACS-sorted adipocytes or fibroblasts from P1 new born mice into adult Rag2^-/- immunodeficient mouse back skin into an excisional wound model. Immunolabelling with anti-Collagen1 or anti-Fibronectin 1 and quantification of associated extracellular matrix in the transplanted regions. g Quantification of adipocyte- and fibroblast-associated ECM in the transplanted regions. n = 3 independent adipocyte samples, n = 6 independent fibroblast or control samples, mean ± SD, unpaired two-tailed t-test. h Representative images and quantification of cathelicidin-related antimicrobial peptide (CRAMP) in GFP positive cells. Data are percentage of CRAMP⁺GFP⁺ cells in total GFP⁺ cells, n = 6 independent samples, mean ± SD, unpaired two-tailed t-test. Arrow heads indicate the wound borders, the stars indicate the examples of double positive staining. Scale bars: c-f, h = 100 µm.