Extended Data Fig. 3: Effect of FAPα cell deletion on leukocyte infiltration.
From: Distinct fibroblast subsets drive inflammation and damage in arthritis
a, Flow cytometry plot of peripheral blood monocytes (numbers, percentage of positive cells) with quantification at day 9 STIA following diphtheria toxin at day 3 and day 5 (n = 6 mice per group). b, Flow cytometry gating strategy for leukocyte populations in digested synovia. c, Numbers of leukocytes and percentage of MHC class II expressing F4/80 cells (M1) in hind limb joints of day 28 persistent STIA mice analysed by flow cytometry (n = 13 mice per group). d, Representative immunohistochemical staining of macrophages (F4/80+, brown; nuclei, blue) in the ankle joint tissue sections at day 12 STIA mice following diphtheria toxin at day 3 and 5 (representative of n = 6 mice). e, Percentage of F4/80+ macrophages staining positive for MHC class II as detected by flow cytometry in day 12 STIA digested synovia from DTR+/− mice following diphtheria toxin at day 3 and 5 (n = 13 mice per group). f, Expression of functional macrophage markers detected by RT−qPCR in sort-purified macrophages (CD45+CD11b+F4/80+) isolated from the synovia of day 12 STIA mice following diphtheria toxin at day 3 and 5 (n = 7 mice per group). g, h, Number of FAPα expressing cells quantified by flow cytometry from digested synovia (n = 8 mice), popliteal (draining) and mesenteric (non-draining) lymph nodes (n = 6 mice) following intra-articular administration of diphtheria toxin into the ankle joint during STIA (harvested day 14) (g) and daily change in weight from baseline (expressed as percentage of original body weight) in STIA mice compared to non-arthritic mice (h) (n = 6 mice). i, Effect of local deletion of synovial FAPα-expressing cells (by intra-articular injection of diphtheria toxin) on ankle joint thickness in the resolving model of STIA model when compared to the wrist joints on the same mouse (non-deleted limbs) (n = 8 mice) and to non-arthritic DTR+ and DTR− injected mice (n = 6 mice per group) with AUC analysis. j, Representative micro-CT images of day 14 STIA and non-arthritic control mice following intra-articular injection of diphtheria toxin and quantification of bone erosion and new bone formation (STIA DTR− n = 10, STIA DTR+ n = 13, DTR− and DTR+ n = 8). k, Quantification of the number of fibroblasts and leukocytes in digested synovia of day 9 STIA mice analysed by flow cytometry following intra-articular administration of diphtheria toxin (n = 8 mice). Statistics: two-way ANOVA with Tukey’s post hoc test (a, c, e, j, k), two-tailed paired Student’s t-test (f, g), one-way ANOVA with Tukey’s multiple comparison tests (i, j). Data are mean ± s.d., except in a, c, e, f, g, k and AUC analysis in i, which are shown as box plots (centre line, median; box limits, upper and lower quartiles; whiskers, maximum and minimum values).
