Pe conjugated anti f4 80 bm8

Single cells suspensions were blocked with Mouse BD Fc Block™ (BD Biosciences, Franklin Lakes, NJ) for 10 min and then stained in FACS buffer (PBS with 4% FBS and 2 mM EDTA) with the following fluorochrome-conjugated antibodies: APC-conjugated anti-CD19 (1D3, #550992, 1:200), APC-Cy7-conjugated anti-CD4 (GK1.5, #552051, 1:200), PE-Cy7-conjugated anti-CD8a (53-6.7, #552877, 1:200), FITC-conjugated anti-CD45 (30-F11, #561088, 1:100), V450-conjugated anti-CD3e (500A2, 560801, 1:100), APC-Cy7-conjugated anti-Ly6C (AL-21, #560596, 1:100), PE-Cy7-conjugated anti-CD45 (30-F11, #552848, 1:100), V450-conjugated anti-CD11c (HL3, #560521, 1:100), PerCP-Cy5.5-conjugated anti-CD11b (M1/70, #561114, 1:100), AF700-conjugated anti-Ly6G (1A8, #561236, 1:100) (from BD Biosciences); PE-conjugated anti-F4/80 (BM8, #12-4801, 1:100), AF700-conjugated anti-CD25 (PC61.5, #56-0251-80, 1:100) (from eBioscience, San Diego, CA). Viability was determined by staining cells with LIVE/DEAD™ Fixable Aqua Dead Cell stain, 1:300 (Thermo Fisher Scientific, Waltham, MA). Flow cytometry was performed on a LSR A flow cytometer (BD Biosciences), and data were analyzed using FlowJo software.