Single cell suspensions were treated with anti-mouse CD16/ CD32 BD Fc Block (2.4G2, Becton Dickinson, NJ, USA) for 10 min followed by staining with appropriate mAbs for 20 minutes on ice. Abs used in this work included PE anti-mPDCA-1clone JF05-1C2.4.1 (Miltenyi Biotec), APC CD11c clone N418 (eBioscience, San Diego, USA), Pacific Blue CD11b clone M1/ 70.15 (Invitrogen), APC-Cy7 B220 clone RA3-6B2 (BD Pharmingen, New York, USA), anti-mouse Ly6G PE-Cy7 (Gr1, eBioscience, San Diego, USA), biotynylated anti-mouse Ly6C (Pharmingen, New York, USA), streptavidin APC-Cy7 (Pharmingen, New York, USA). Flow cytometric analysis and sorting was performed using LSRII (Becton Dickinson, NJ, USA. The data were analyzed using FACSDiva (Becton Dickinson) software.