Extended Data Fig. 3: Mouse single-cell landscape in joints and skin.
(a) Graphical cartoon showing the strategy to cell sort target populations used for scRNA-seq. Target populations were individually hash-tagged before generating the scRNA-seq library. (b) UMAP plot of the hash-tag identified ankle joint cells in the scRNA-seq dataset from Kaedetg mice (BALB/c and C57BL/6) on day 7 of the IL-23OE model. Refers to Fig. 2a. Detailed annotation with Seurat-identified clusters shown. (c) CellRank identification of terminal (left) and initial states (right) in KaedeRED myeloid cells. (d) Coarse-grained matrix of transition probabilities between states calculated by CellRank. (e) Significantly enriched GO-BP terms in Ly6c2+ monocyte cluster, when compared to all other cell types in the joint. Selection criteria: BH-adjusted P value < 0.05. (f) UMAP plot of the hash-tag identified skin leukocytes in the scRNA-seq dataset from Kaedetg mice (BALB/c 2,494 cells, N = 1 and C57BL/6 3,024 cells, N = 1) on day 7 of the IL-23OE model. Annotated Seurat identified clusters shown (left). KaedeGREEN and KaedeRED cells are highlighted (right). (g) Significantly enriched GO-BP terms in skin CD2+MHC-II+CCR2+ myeloid precursors (PCs) cluster, when compared to other cell types. Selection criteria: BH-adjusted P value < 0.05. (h) Representative flow cytometry plots showing the gating strategy for the identification of CD2+MHC-II+CCR2+ skin-derived PCs in the joint. Red gate on T/B lymphocytes as internal control for CD2 and MHC-II. (i) Quantification of CD2+MHC-II+ myeloid cells in the skin of BALB/c animals after IL-23OE over time. Graph shows median, quartiles, min-max.; N = 5, per timepoint and condition; P-values calculated by one-way ANOVA with Tukey´s post hoc test. (j, k) Representative flow cytometry plots showing the gating of CD2+MHC-II+ skin-derived myeloid PCs in the bone marrow (j, left) or small intestine (k, left) and their respective quantification after IL-23OE on day 7 in BALB/c and C57BL/6 mice (j, k right). Graph (j) shows median, quartiles, min-max.; N = 5, per condition; P values calculated by one-way ANOVA with Tukey’s post hoc test. Graph (k) shows median, min-max.; N = 2, per condition; no P values calculated.
