Labeling, enrichment and functional blockade of Ly6Chi monocytes

Mice received 200 μL clodronate liposomes intravenously and 20 h later, 0.25 mL of 0.5 μm Fluoresbrite fluorescein isothiocyanate (FITC)-dyed (YG) plain microspheres (Polysciences, USA) were injected intravenously to selectively label activated monocytes. To specifically enrich Ly6C^hi cells, mice were injected intravenously with 0.25 mL clodronate liposomes (Formumax, USA) or PBS liposomes, and the peripheral monocyte pool was replenished within 24 h. For the functional blockade of Ly6C^hi monocytes, mice were intraperitoneally administered with anti-mouse Ly6C antibody (Clone HK 1.4, 100 μg) or isotyoe controls (Rat IgG2c, κ isotype control, 100 μg).