Anti ccr7 apc cy7

LNMCs and PBMCs were stained with anti-CCR7–APC-Cy7 (G043H7, BioLegend) for 10 minutes at 37 °C. Cells were then labeled with LIVE/DEAD Fixable Aqua (Thermo Fisher Scientific) for 10 minutes at room temperature to identify non-viable events, stained with a cocktail of directly-conjugated monoclonal antibodies for 20 minutes at room temperature to detect surface markers, fixed/permeabilized using a Cytofix/Cytoperm Kit (BD Biosciences), stained with another cocktail of directly-conjugated monoclonal antibodies for 1 hour at room temperature to detect intracellular markers, and fixed in 1% paraformaldehyde (Sigma-Aldrich). Data were acquired using an LSRII flow cytometer (BD Biosciences) and analyzed with FlowJo software v9.9.4 (Tree Star Inc.).