Journal: Nature Communications
Article Title: Single-cell transcriptomics reveals aberrant skin-resident cell populations and identifies fibroblasts as a determinant in rosacea
doi: 10.1038/s41467-024-52946-7
Figure Lengend Snippet: a UMAP plots showing keratinocyte subclusters and sample conditions. b Bar graph showing subpopulation percentage. c Feature plot showing CD74 expression in keratinocytes. d The percentage of CD74 + keratinocytes ( n = 3 samples for each group). e Immunohistochemistry of KRT14, CD74 in HS, normal skin of patients (NS), and lesional skin of ETR, PPR, PhR. White arrows indicate CD74 + keratinocytes. Scale bar, 50 μm. f Quantification of CD74 + keratinocytes ( n = 6/6/6/6/5 samples for HS/NS/ETR/PPR/PhR group used in e ). g , h Top-ranked enriched pathways in CD74 + keratinocytes of L versus HS ( g ), L versus N ( h ). NES indicates enrichment scores. The color keys from red to blue indicate the P -value range. Two-sided permutation test without multiple comparison adjustments was used. i Heatmap of skin barrier-related genes in CD74 + keratinocytes. j Heatmap of IFN-related genes in CD74 + keratinocytes. k Immunohistochemistry of IRF1, KRT14. Scale bar, 100 μm. l Quantification of IRF1 + keratinocytes in epidermis n = 6/6/6/6/5 samples for HS/NS/ETR/PPR/PhR group used in k ). m The back skins of IgG and anti-IFNγ antibody-treated mice intradermally injected with LL37 or control vehicle. Images were acquired 48 h after the first LL37 injection. Below panels, magnified images of yellow boxed areas. n The severity of the rosacea-like symptoms was determined with the redness area and score ( n = 6). o Dermal infiltrating cells were quantified ( n = 6). p The relative mRNA levels of Il1β, Il6 , Tnfα in mice skins ( n = 6). q The relative mRNA levels of Cldn4 , Cldn10 , Cldn23 in mice skins ( n = 6). r Immunohistochemistry of CLDN4, KRT14 in mice skins. Scale bar, 50 μm. Epi, Epidermis. Der, Dermis. s Quantification of relative fluorescence intensity for CLDN4 in epidermis ( n = 6). t Quantification of TEWL ( n = 6). All results are representative of at least three independent experiments. Data are presented as mean ± SEM, and P -values were determined by one-way ANOVA with Tukey’s post hoc test ( f , l , n , o , p , q , s , t ) and two-tailed unpaired (L vs HS) or paired (L vs N) Student’s t -test ( d ).
Article Snippet: The following primary antibodies were used: Goat anti-PDGFRA antibody (1:100, AF1062, R&D systems), Rabbit anti-PTGDS antibody (1:200, 10004344, Cayman), Rabbit anti-PDGFRA (1:500, ab203491, abcam), Goat anti-CCL19 (1:100, AF880, R&D systems), Rabbit anti-CD4 (1:500, ab183685, abcam), Rat anti-CCR7 (1:100, MAB3477, R&D systems), Rat anti-IFNγ (1:100, 14-7311-81, Thermo), Rabbit anti-MBP (1:1500, 78896, Cell signaling), Sheep anti-CD74 (1:100, AF7478, R&D systems), Rat anti-F4/80 (1:100, 14-4801-81, Thermo), Goat anti-CXCL10 antibody (1:100, AF466, R&D systems), Rabbit anti-MLC2 (1:400, 15354-1-AP, Proteintech), Mouse anti-α-SMA (1:5000, ab7817, abcam).
Techniques: Expressing, Immunohistochemistry, Comparison, Injection, Control, Fluorescence, Two Tailed Test