Article Title: Mechanisms of stretch-mediated skin expansion at single-cell resolution
Figure Lengend Snippet: Genetic signature of TPA treated and expanded epidermis. a , Representative FACS plots showing the strategy used to isolate basal cells. Single living cells were gated by debris exclusion (P1), DAPI exclusion (P2), doublet elimination (P3) and basal IFE Integrin-α6 high CD34 neg cell were sorted (P4). n=10 independent experiments. b, c , mRNA expression of genes that were upregulated in basal cells at EXP D4 (n = 3) and in cells treated with TPA (n = 2). These genes are related to a generic stress signature (b) , regulating ECM remodeling and cytoskeleton, important for cell survival and cell cycle (c). Bars are mean with s.e.m. d , Representative images of Paxillin immunostaining color-coded for the signal intensity with ImageJ. Protein expression is visualized as a color gradient going from black to yellow with black as indicator of no expression and yellow as indicator of maximal expression. Scale bar, 10 μm e , Quantification of the average integrated density signal for Paxillin as in d. Each data point is the average of 3 sections per mouse (n = 3 mice per condition). f , Geometric mean fluorescence intensity for the indicated integrin in CTRL (grey, n = 4 mice) and EXP D4 (red, n = 6 mice) from FACS analysis of basal IFE Integrin-α6 high CD34 neg cells. g-k , ATAC-seq profiles showing increasing accessibility of chromatin regions that are specifically remodeled during mechanical expansion (CTRL in grey and EXP D2 in orange). l , Quantification of the number of cells FOSL1+ in the basal layer related to . m , Immunostaining on skin sections for c-JUN (white) in control and EXP D4. n , Quantification of the number of cells c-JUN+ in the basal layer related to m. o . p . q , Immunohistochemistry on paraffin sections for c-FOS in control and EXPD4. Scale bar, 20 μm. r , Quantification of the number of cells c-FOSL+ in the basal layer related to q. s , Immunofluorescence on tissue sections for pSTAT3 in green and K14 (red) to identified the epidermis. Scale bar, 20 μm. t , Quantification of the number of cells positive for pSTAT3 in the basal layer related to s. l, n, o, p, r, t , 3 sections quantified per n = number of mice and total number of cells indicated in parentheses d, m, q, s , Dashed lines delineate the basal lamina. e, f, l, n, o, p, r, t , Two-tailed Mann–Whitney test, Mean + s.e.m. Fig. 3d
Article Snippet: Basal IFE and infundibulum cells were stained using PEconjugated anti-α6-integrin (clone GoH3, 1:200, ebioscience) or FITC- conjugated anti-α6-integrin (clone GoH3, 1:200, ebioscience) and bulge cells were stained with biotinylated CD34 (clone RAM34; 1:50, BD Biosciences).
Techniques: FACS, Expressing, Immunostaining, Mouse Assay, Fluorescence, Immunohistochemistry, Immunofluorescence, Two Tailed Test, MANN-WHITNEY