Cells from spleen, liver and BM were isolated as previously described (28 (link)). Antibodies are listed in Supplemental Figure 1 . Samples were acquired using LSR Fortessa cytometer (BD Biosciences) and BD FACSDiva software (v.8.0.1, BD Biosciences) and analyzed with FlowJo software (Tree Star). Cell sorting was performed using FACSAria III (BD Biosciences). For the evaluation of IFN-γ expression, cells were left untreated or stimulated with PMA/Ionomycin (Sigma-Aldrich) for 2 h or IL-2 (1,000 U/ml, Hoffmann-La Roche Inc.) and IL-12 (10 ng/ml), or IL-12 (10 ng/ml) and IL-18 (100 ng/ml) (R&D Systems) for 6 h (with the addition of GolgiPlug, from BD Biosciences).
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