Anti mouse tim 3 pe

The isolated immune cells were resuspended in 0.1 mM PBS and passed through a 70-μm strainer (BD Biosciences, NJ,USA). Samples were analyzed with the following antibodies: anti-human CD14 FITC (325604, Biolegend, San Diego, CA, USA); anti-human Tim-3 PE (345006, Biolegend, San Diego, CA, USA); anti-mouse CD45 APC-eFlour780 (47-0451-82, eBioscience, San Diego, CA, USA), anti-mouse F4/80 PE-eFlour610 (61-4801-82, eBioscience, San Diego, CA, USA); anti-mouse CD11b PE-Cy7 (25-0112-82, eBioscience, San Diego, CA, USA), CD3 APC (100236, Biolegend, San Diego, CA, USA), CD4 percycy5.5 (103132, Biolegend, San Diego, CA, USA), CD8 FITC (100706, Biolegend, San Diego, CA, USA), CD11c APC (17-0114-81, eBioscience, San Diego, CA, USA), NK1.1 pecy7 (25-5941-82, eBioscience, San Diego, CA, USA), anti-mouse Tim-3 PE (12-5870-82, eBioscience, San Diego, CA, USA). Antibodies and their isotype-matched negative control antibodies were incubated with cells at 4 °C for 30 min in dark. Cells were washed with 0.1 mM PBS. The samples were subjected and detected by a Beckman CytoFLEX FCM, and the data were analyzed by CytExpert 2.0 software.