Extended Data Fig. 3: Fate mapping of synovial lining macrophages during arthritis.

a, Gating strategy for CD45⁺CD11b⁺Ly6G⁻CD115⁺ classical Ly6C^high and non-classical Ly6C^low monocytes of Cx3cr1^creERR26-tdTomato mice. b, Gating strategy for DAPI⁻ living, CD45⁺CD11b⁺Ly6G⁻F4/80⁺ macrophages of Cx3cr1^creERR26-tdTomato mice. c, Evaluation of tdTomato expression in blood monocytes and synovial macrophages two days and four weeks after tamoxifen pulse. Mean ± s.e.m.; n = 4 per group. d, e, BFM images of knee joint synovial membranes of Cx3cr1^creERtdTomato mice four weeks after systemic tamoxifen pulse (d) and five days after local injection of (Z)-4-hydroxytamoxifen (e) at day 0 and day 7 after the induction of K/BxN STA showing selective tdTomato (red) expression in synovial lining macrophages. The smaller graphs to the right in e show the absence of tdTomato expression in blood monocytes after local (Z)-4-hydroxytamoxifen injection. Scale bars, 25 µm. f, Gating strategy for DAPI⁻ living, CD45⁺CD11b⁺Ly6G⁻F4/80⁺ macrophages of Cx3cr1^creERR26-tdTomato mice four weeks after tamoxifen pulse, used to calculate the absolute numbers of tdTomato⁺ macrophages during steady state and K/BxN STA. g, Gating strategy after EdU labelling of proliferating macrophages (CD45⁺CD11b⁺Ly6G⁻ F4/80⁺) of Cx3cr1^creERR26-tdTomato mice. h, Quantification of total tdTomato⁺ and tdTomato⁻ macrophages in paws of Cx3cr1^creERR26-tdTomato mice 4 weeks upon tamoxifen pulse at day 0, 2 and 5 after induction of STA. Mean ± s.e.m. Day 0, n = 6; day 2, n = 5; day 5, n = 6.

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