Wild-type and BMPR1αT− mice were immunized with 100 μg of LCMV peptide gp61-80 (AnaSpec) in CFA in the footpads. After 1 month, a booster dose was applied, and mice were sacrificed 7 days later. Draining (popliteal) lymph nodes and spleen cells (2 × 105 cells per well) were isolated, and cytokine-secreting cells were quantified using ELISPOT assay and ELISPOT Ready-SET-Go kit reagents (eBioscience). Cytokines were detected using antibodies specific for IFN-γ (capture, R4-6A2; detection, XMG1.2), IL-4 (capture, 11B11; detection, BVD6-24G2), IL-17 (capture, eBio17CK15A5; detection, eBio17B7), IL-2 (capture, JES6-1A12; detection, JES6-5H4), and IL-10 (capture, JES5-16E3; detection, JES5-2A5), and spots were quantified with ImmunoSpot software (CTL) on an S6 Macro reader.

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