Extended Data Fig. 4: Analysis of dermal DCs and wound healing.
From: Targeting SLC7A11 improves efferocytosis by dendritic cells and wound healing in diabetes
a, Gating strategy of enriched phagocytes after digestion of ears and depletion of lymphocytes. b, Immunofluorescent images of skin sections from unwounded PDGFR-GFP mice depicting Slc7a11-positive (red) cells. Nuclei were stained with DAPI. Scale bar: 50 μm. c, Annotations of innate immune cell populations arising in lesional skin. d, Frequencies of Slc7a11 expression in innate immune cells of lesional and non-lesional skin. e, Representative images of wounds of mice treated with erastin or vehicle at day 10 post-wounding. f, Wound healing dynamics comparing WT mice after full-thickness wounding and a single topical administration of apoptotic targets at the day of wounding or erastin and vehicle only at day 0 till day 2 (n = 8 per group). Data represent means ± SEM. g, Comparison of wound closure at day 2 post wounding in WT mice treated with RSL3 regimen (n = 9) versus erastin regimen (n = 7) or DMSO regimen (n = 7). Data represent means ± SEM.* P < 0.05; ** P < 0.01. One-way ANOVA with Tukey’s multiple comparisons test. Box and whiskers show minimum to maximum values with all independent replicates, center denotes median. h, i, Erastin regimen promotes in vivo migration of keratinocytes during wound healing. PDGFRa-H2BeGFP mice were treated with erastin or DMSO regimen after full-thickness wounding with an 8 mm punch biopsy. Skin sections at day 4 post-wounding were stained with h, Itga5 (n = 4 per condition, keratinocytes; n = 4 DMSO-fibroblasts; n = 7 Erastin-fibroblasts) or i, Ki67 (n = 4 per condition, keratinocytes; n = 4 DMSO-fibroblasts; n = 5 Erastin-fibroblasts) Quantification (right) and representative immunofluorescent images (left) of skin sections with respective treatments. Nuclei were stained with DAPI (blue). Violins plots show minimum to maximum values with all independent replicates, centre denotes median. (*P = 0.0420; unpaired two-tailed t-test); ns:not-significant. Scale bar: 150 μm. j, Percentage of scratch wound closure (re-epithelialization) of mouse primary Slc7a11 WT (n = 5) and KO (n = 4) keratinocytes pretreated with mitomycin C. Slc7a11KO keratinocytes show no difference indicating that the effect on migration in vivo (h) is not cell intrinsic. Data are expressed as mean ± SD.
