Fig 2: Changes in cell numbers for cell populations in unwounded skin and WIHN wounds. Cells were isolated from digested unwounded skin (pw0) or at different pw along wound healing, stained with antibodies against cell surface markers or intracellular FOXP3 and analyzed with flow cytometry. (A) Composition of skin (pw0) or wounds (other pw) in terms of dermal compartment, immune cells and epidermis. (B) Changes in epithelial cell number expressed as cell/1 × 103 CHD1+ cells. (C) Changes in suprabasal and basal Bul. (D) Changes in numbers for fib expressed as cell/1 × 103 CHD1−/PTPRC− cells. (E) Changes in DP and endothelial cells expressed as cell/1 × 103 CHD1−/PTPRC− cells. (F) Changes in numbers for DC, mono and neutrophil expressed as cell/1 × 103 PTPRC+ cells. (G) Changes in CD206(MRC1) + and − cells expressed as cell/1 × 103 PTPRC+ cells. (H) Changes in numbers for T cells expressed as cell/1 × 103 PTPRC+/CD3+ cells. (I) Exemplary dot plot of CD4 and CD8 expression of CD3+/PTPRC+ cells at pw9. (J) Changes in numbers for δγT cells expressed as cell/1 × 103 PTPRC+/CD3+ cells and Treg expressed as cell/1 × 103 PTPRC+/CD3+/CD4+ cells. Results are expressed as mean ± SEM, n = 3–8. Comparison between pw0 and subsequent pw values was carried out individually with a t-Test, p value = * ≤ 0.05, ** ≤ 0.01.

Fig 3: Treg-specific deletion of Il1r2 led to increased antitumor efficacy of ICI therapy. (A and C) MC38 cells (2×106) were inoculated i.d. into the right flank of Foxp3Cre and Il1r2fl/flFoxp3Cre male mice. αPD-1 (A) or αCTLA-4 (C) mAbs, and IgG controls were used in the treatment on day 5 after tumor inoculation and again every 4 days for four times. Tumor sizes were monitored every 2 days. (B and D) The overall survival of different groups was monitored in two-group mice challenging with αPD-1 or αCTLA-4 mAbs and control IgG, respectively. (A and B: n=8 in each subgroup. C and D: n=6 in each subgroup). (E) Another experiment was set up using female mice (6 weeks old, n=10), and after three times of αPD-1 or IgG treatment, the tumor tissues were collected, and the TILs were analyzed by using FACS. The representative flow cytometry plots showed the percentages of CD4+ TILs, CD8+ TILs and Tregs in different group mice. (F) Statistical analysis of the percentages of total CD45+ TILs, CD4+ TILs in total CD45+ TILs, CD8+ TILs in total CD45+ TILs, Treg in CD4+ TILs in different group mice. Data represent at least three independent experiments. *P<0.05, **p<0.01, ***p<0.001 and ****p<0.0001 by Student’s t-test, two-way analysis of variance, or log-rank survival analysis. ICI, immune checkpoint inhibitor; TILs, tumor-infiltrating lymphocytes; Treg, regulatory T cell.

Fig 4: Xiao-Yin-Fang lessened imiquimod-induced psoriasis-like dermatitis on mouse ears. (A) Schematic of the experimental procedures. Mice were given XYF or H2O by gavage twice daily from day 0 to day 9, and received IMQ or VAS application on mouse ears once a day from day 5 to day 9 (B) Representative pictures of mouse ear lesions. (C, D) Evaluation of PASI score (C) and skin swelling (D) of mouse ears (n = 27, three independent experiments) (E) H&E staining and calculated epidermal acanthosis (F) (x200; bar = 50 μm; n = 18, three independent experiments). (G) Ki-67 staining and assessment of its H-score of epidermal fields (H) (x200; bar = 50 μm; n = 18, three independent experiments). (I–K) Freshly isolated dermal cells were obtained and stained with anti-CD45, MHC-II, CD11b, F4/80, CD11c, Ly6C, and Ly6G antibodies, which were analyzed by flow cytometry. (I) Representative scatter plots of dermal neutrophils and inflammatory monocytes. The ratios and cell numbers of dermal neutrophils (J) and inflammatory monocytes (K) (n = 16, three independent experiments). The data were presented as mean ± s.e.m.

Fig 5: Dynamic Features of Tracheal Myeloid Populations after Injury(A) UMAP visualization of all SCRNA-seq data across airway injury and repair overlaid (3 animals pooled into 1 sample for each); captured cells: ctrl = 458, 1 dpP = 1,625, 4 dpP = 1,507, 7 dpP = 1,717.(B) UMAP visualization of SCRNA-seq data on indicated days post-injury.(C) Violin plots of M2-like macrophage gene signature, Arg1 and Arg2.(D) Violin plots of classic monocyte/macrophage gene signature, Ccr2 and Cx3cr1.(E) Violin plots of cell cycle genes associated with G2/M and S phase.(F) Representative FACS plot and quantification of YARG expression gated on CD45+;F4/80+ (n = 3 mice per time point).(G) FACS analysis and quantification of Ccr2-CreERT2;ROSA-tdTom+ cells from YARG animals after tamoxifen administration and sham or polidocanol treatment (n = 3, mice per time point).(H) Immunofluorescence analysis of Ccr2-CreERT2;ROSA-tdTom animals; Krt5 (basal cells, green), Ccr2-CreERT2;ROSA-tdTom lineage trace (red), and Höechst nuclear stain (blue); scale bar, 15 μm.(I) Histology of wild-type animals; Krt5 (red), CD45 (green), nuclear stain Höechst, scale bar, 15 μm.(J) Confocal image of peeled epithelial sheet showing F4/80+ myeloid cells (green) and Krt5+ epithelial basal cells (red), Höechst nuclear stain. See also Figure S2 and Tables S2 and S4.