Poly 1 poly c

For cell cycle analysis, EdU and BrdU were used. Both are incorporated during DNA synthesis and, combined, allow analysis of cell cycle kinetics^{45 (link)}. Pulse-chase timings were optimised to achieve consistent in vivo incorporation of EdU and BrdU, and later detection in the cell populations of interest. EdU (1 mg/mouse) and BrdU (2 mg/mouse) were administered via tail vein injection 2 h apart, unless otherwise stated; 30 min later, mice were culled and the tissue was isolated. Poly-I-poly-C (Sigma) was administered intraperitoneally as a single dose of 100 µg, 24 h prior to culling.

Human IL-1α expression was induced in 8-week-old male human IL-1α conditional transgenic mice (hIL-1 cTg) by injecting 200 µl of a solution containing 250 µg of PolyI-PolyC (Sigma-Aldrich Co., St. Louis, MO, USA) for 3 consecutive days intraperitoneally. Some mice were induced with CD-4-depletive or ISO type control antibody (each 5 mg/kg)^{28 (link)}, followed by additional PolyI-PolyC injection at 9 and 10 weeks of age. Some hIL-1 cTg mice were not treated with PolyI-PolyC. Arthritis severity was evaluated by measuring the ankle thickness before and after PolyI-PolyC injection at various time points.

C57BL/6 mice were maintained under specific-pathogen-free (SPF) conditions. Male mice aged 8 to 10 weeks were subjected to sublethal (600 cGy) total body irradiation (TBI) with γ-radiation from a ⁶⁰Co irradiator. The mice were injected intraperitoneally (i.p.) with mD1R (4 mg/kg)19 (link) or phosphate-buffered saline (PBS) 2 h post irradiation, and this was followed by daily injection of the same reagent for 7 or 14 days. In some experiments, male mice were injected i.p. with CTX (150 mg/kg) (Sigma-Aldrich, St Louis, MO), followed by treatment with mD1R or PBS as above. RBP-J-floxed (RBP-J^f) mice were described previously20 (link). For the induction of RBP-J deletion, RBP-J^f/f mice were mated with Mx-Cre mice, and mice with suitable genotypes were injected with poly(I)-poly(C) (Sigma-Aldrich) as described previously20 (link). To monitor cell proliferation, the mice were injected i.p. with 5-bromo-2-deoxyuridine (BrdU; Sigma-Aldrich) (100 mg/kg in PBS) every two days after irradiation and were maintained with drinking water containing 1 mg/ml BrdU for 14 days. Granulocyte colony-stimulating factor (G-CSF) was administered i.p. at a dose of 50 μg/kg. All animal experiments were approved by and performed in accordance with guidelines from the Animal Experiment Administration Committee of the Fourth Military Medical University.