Fig. 4: Joint fibroblasts prime migrating precursors to proinflammatory or anti-inflammatory phenotypes.
a, TradeSeq-fitted smooth expression of UCell scores of genes dynamically associated with either BALB/c (top) or C57BL/6 (bottom) mice over pseudotime from Fig. 2e in the scRNA-seq dataset of ankle joints from Kaedetg mice (BALB/c and C57BL/6 strains) on day 7 of the IL-23OE model; Padj, adjusted P value. b, UCell scores for GO-BP terms GO:0050728 (anti-inflammatory response) and GO:0050729 (proinflammatory response) on the phagocyte compartment from Fig. 2a stratified by stain and photoconversion status. P values were calculated with a two-sided Wilcoxon signed-rank test and adjusted for multiple comparisons using the BH method. c, Circle plot showing the communication probability between myeloid cells (KaedeGREEN or KaedeRED) and other cell types in the joints associated with either BALB/c (top) or C57BL/6 (bottom) mice. Line thickness corresponds to communication probability determined using CellChat; ECs, endothelial cells; Mφ, macrophage. d, UMAP plot of Seurat-identified subclusters among fibroblasts in the scRNA-seq dataset of ankle joints from Kaedetg mice (BALB/c and C57BL/6 strains) on day 7 of the IL-23OE model. e, Expression of the most relevant marker genes among each fibroblast cluster identified in d. f, Comparison of UCell scores of the C57BL/6-associated gene signature (fibroblast-only differentially expressed genes in C57BL/6 mice compared to BALB/c mice) between different subclusters of joint fibroblasts; DEG, differentially expressed gene. g, C57BL/6-associated relative likelihood of different subclusters of fibroblasts determined by MELD. h, Quantification of CD200+ fibroblasts in the joints of BALB/c and C57BL/6 mice on day 7. The graph shows the median, quartiles and minimum–maximum; N = 4 per time point and condition. P values were calculated by one-way ANOVA with a Tukey’s post hoc test. i, Quantification of CD200+ fibroblasts in the joints of BALB/c mice on days 2, 4 and 7. The graph shows the median, quartiles and minimum–maximum; N = 4 per time point and condition. P values were calculated by one-way ANOVA with a Tukey’s post hoc test. j, Expression of Cd200r1 among the different cell clusters in the scRNA-seq dataset of ankle joints from Kaedetg mice (BALB/c and C57BL/6 strains) on day 7 of the IL-23OE model. k, Heat map of median fold change gene expression in macrophage–fibroblast cocultures treated with anti-CD200 antagonist (ant.; OX-90) or isotype control; N = 7; FC, fold change. l, Representative images of MRI scans and micrographs of H&E-stained ankles of C57BL/6 animals at day 21 after IL-23OE with or without anti-CD200 antagonist (OX-90) treatment. MRI quantification of arthritis at day 21 is shown on the right. Arrowheads indicate inflammation, and stars indicate the talar bone. The graph shows the median, quartiles and minimum–maximum; N = 4 per condition. P values were calculated by two-sided Mann–Whitney U-test. m, Representative images of MRI scans and micrographs of H&E-stained ankles of BALB/c animals at day 21 after IL-23OE with or without anti-CD200R1 agonist (ago.; OX-110) treatment. MRI quantification of arthritis at day 21 is shown on the right. Arrowheads indicate inflammation, and stars indicate the talar bone. The graph shows the median, quartiles and minimum–maximum; N = 4 per condition. P values were calculated by two-sided Mann–Whitney U-test.
