IMQ model and in vivo treatments

Cream containing 5% IMQ (Aldara; 3M Pharmaceuticals) was topically applied to the dorsal and ventral sides of both ears. Treatment was repeated with 20 mg of cream per ear every 24 h for 7 consecutive days. Mice were sacrificed 24 h after the last application. Ear thickness was measured before first application and daily before treatment with a dial thickness gauge (Mitutoyo UK). The mean of two measurements (one per ear) was determined. The following unconjugated monoclonal antibodies were administered in vivo as described in detail in every figure; control groups received suitable control IgG via the same route and at the same dose. To deplete Foxp3⁺ cells in B6 Foxp3^hCD2, 250 µg anti-human CD2 (YTH 655) was administered in B6 Foxp3^hCD2 mice i.v. on days −1 and 0, and i.p. on days 2, 4, and 6 of IMQ treatment. Anti-mouse CD8 antibody (53–6.72) was injected i.p. at 120 µg/dose on days −1 and 0, and on days 2, 4, and 6 of IMQ treatment. MNPs were depleted using clodronate liposomes. 0.2 ml of clodronate or PBS (control) liposomes were injected i.p. on days −1, 0, 2, 4, and 6 12 h after anti-hCD2 injection. To block IFN-I signaling, mice were treated with 1 mg of anti-mouse IFNAR1 antibody (MAR1-5A3) i.p. on day −1 of IMQ treatment.