At day 8 of CFA, paw tissue was minced and digested in Hanks’ balanced salt solution (HBSS) containing collagenase I (1 mg/ml; Roche) and dispase (2.4 U/ml; Gibco) for 90 min at 37°C under agitation. After digestion, cells were filtered through a 70-mm mesh and washed in PBS 1% FBS. After blocking with CD16/CD32 (1/100 Fc Block, eBioscience) on ice for 15 min, cells were stained for 20 min with an anti-mouse CD3–fluorescein isothiocyanate (clone 145-2C11, BD Biosciences) and anti-mouse CD11b-PercP Cy5.5 (clone M1/70, eBioscience) and then analyzed on a BD FACSCanto II (BD Biosciences).

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