α gr1 clone rb6 8c5

For flow cytometry analysis, isolated cells (from cNTs, lymph nodes, or spleens) were aliquoted at 500,000 cells per 5 ml tube, and pre-treated with Fc block (hybridoma clone 2.4G2) prior to cell staining. Staining was done in panels using the following antibodies: α-CD3-APC (clone 145-2C11), α-CD3-PE Cy7 (clone 53–7.3), α-CD8 PE Cy7 (clone 53–6.7), α-CD45 PE (clone 30-F11), α-CD19 (clone MB19-1), α-NK1.1 (clone PK136), α-CD11c (clone N418), α-F4/80 (clone BM8), and α-GR1 (clone RB6-8C5) (all from eBioScience); α-CD4 APC Cy7 (clone GK1.5; Biolegend); and α-CD45 Alexafluor 700 (clone 30-F11; BD Biosciences). Dead cells were excluded using 7-AAD (BD Biosciences). Antibodies to stain cells for each panel were added, mixed, and incubated on ice in the dark for 30 minutes. Cells were washed twice with 1× PBS +5% FBS and resuspended in 500 µl of 1× PBS +5% FBS. Stained cells were run on a BD FACS Canto II flow cytometer (BD Biosciences). Standard compensations were used for each tissue using FACSdiva software.