Anti mouse cd62l

Flow cytometry was performed as previously described (33 ). Briefly, single-cell suspensions were isolated from spleens or lymph nodes by crushing with forceps, or from sciatic nerves by mincing and digestion in 2 mg/ml collagenase. Cells were stained with live/dead fixable yellow dye (Life Technologies), anti-mouse CD4 (Biolegend clone GK1.5), anti-mouse CD44 (eBioscience clone IM7), anti-mouse CD62L (eBioscience clone MEL-14), anti-mouse S1PR1 (R&D clone 713412), anti-mouse IFN-γ (eBioscience clone XMG1.2), and anti-mouse IL-10 (BD clone JES5-16E3) antibodies. For intracellular cytokine staining, cells were stimulated with PMA/ionomycin for 4 hours at 37°C 5% CO₂, and permeabilized using BD Cytofix/Cytoperm according to manufacturer’s instructions. Cells were analyzed on a CyAn ADP Analyzer (Beckman Coulter). Data were analyzed using FlowJo X.

Anti–mouse CD4, anti–mouse T-cell receptor (TCR) β, Anti–mouse CD44, anti–mouse CD62L, anti-mouse CD25, anti–mouse Foxp3, anti-CD8, and anti–B220 antibodies were purchased from eBio-science (San Diego, CA). Anti-mouse CD138 antibody was purchased from BD Biosciences (Franklin Lakes, NJ).