Fig. 1. Phenotype of the fibroblast subpopulations. (a) (i) Schematics of the procedure followed for the isolation of papillary and reticular fibroblasts and (ii) the representative gating strategy of the sorting. After negative selection for CD324⁻, CD45⁻ and CD31⁻ and positive selection for CD90⁺, two populations were FACS sorted – CD39⁺CD26⁻ cells corresponded to the papillary fibroblasts, while CD36⁺ cells to the reticular fraction. (iii) Percentage of isolated fibroblasts after sorting. (b) (i) Representative images of the F-actin stained cytoskeleton and respective quantification of the area of the cells. Scale bar = 50 μm. (ii) Representative immunocytochemistry images of the expression of podoplanin (PDNP), transglutaminase-2 (TGM2) and fibroblast activated protein (FAP) after isolation. Scale bar = 100 μm. Cell nuclei were counterstained with DAPI. Individual channel images are presented in ESI Fig. 2.† (c) Phenotype of the papillary and reticular fibroblasts after passage 1 (P1) and 2 (P2) determined by flow cytometry and respective representation of their proliferation as per DNA quantification along the culture time. Quantitative results are expressed as the mean ± standard deviation where n = 3, * p < 0.05, in the two-tailed unpaired Mann–Whitney test.