The reagents that have been used in this study were supplied by Sigma‐Aldrich (Saint‐Quentin Fallavier, France). Clobetasol was chosen as it is a potent topical steroid widely used in dermatology for various skin diseases that is able to induce skin atrophy and healing delay.
The following antibodies were used for immunofluorescence staining: rat anti‐mouse F4.80 (1:250; Abcam, Cambridge, UK), rat anti‐Gr‐1 (1:250; eBioscience, Villebon‐sur‐Yvette, France), rat anti‐CD3 (1:250; Biolegend, Paris, France), and rat anti‐mouse CD31 (1:200; BD Biosciences, Le Pont de Claix, France), rabbit anti‐K14 (1:500; Biolegend). The sections were afterwards counterstained with 4,6‐diamidino‐2‐phenylindole 0.3 μg·ml−1 (Sigma‐Aldrich, Saint‐Quentin‐Fallavier, France). Flow cytometry analysis was performed using PE/Cy7‐conjugated rat anti‐F4/80 (1:200, eBioscience), FITC‐conjugated rat anti‐CD31 (1:100, Biolegend), APC‐conjugated rat anti‐CD45 (1:1000; BD Bioscience), APC/Cy7‐conjugated rat anti CD11b (1:100; Biolegend), PE‐conjugated rat anti‐Ly6C (1:100, Biolegend), and FITC‐conjugated rat anti‐CD206 (1:100; Biolegend). The immuno‐related procedures used comply with the recommendations made by the British Journal of Pharmacology (Alexander et al., 2018 (link)).
The following antibodies were used for immunofluorescence staining: rat anti‐mouse F4.80 (1:250; Abcam, Cambridge, UK), rat anti‐Gr‐1 (1:250; eBioscience, Villebon‐sur‐Yvette, France), rat anti‐CD3 (1:250; Biolegend, Paris, France), and rat anti‐mouse CD31 (1:200; BD Biosciences, Le Pont de Claix, France), rabbit anti‐K14 (1:500; Biolegend). The sections were afterwards counterstained with 4,6‐diamidino‐2‐phenylindole 0.3 μg·ml−1 (Sigma‐Aldrich, Saint‐Quentin‐Fallavier, France). Flow cytometry analysis was performed using PE/Cy7‐conjugated rat anti‐F4/80 (1:200, eBioscience), FITC‐conjugated rat anti‐CD31 (1:100, Biolegend), APC‐conjugated rat anti‐CD45 (1:1000; BD Bioscience), APC/Cy7‐conjugated rat anti CD11b (1:100; Biolegend), PE‐conjugated rat anti‐Ly6C (1:100, Biolegend), and FITC‐conjugated rat anti‐CD206 (1:100; Biolegend). The immuno‐related procedures used comply with the recommendations made by the British Journal of Pharmacology (Alexander et al., 2018 (link)).