Percp cyanine5.5 anti mouse f4 80

CpG ODN 2359 was obtained from Tsingke Biological Technology Co. (Beijing, China). Anti-OX40 agonistic antibody (αOX40, Clone OX-86) was provided by GenScript USA Inc. Murine anti-PD-1 antibody (G4C2) was provided by Shanghai Junshi Biosciences Co.,Ltd (Suzhou, China).Monoclonal antibodies (mAbs) used for flow cytometry were listed as flow: FITC anti-mouse CD45 (Biolegend, USA), PE/Dazzle™ anti-mouse CD3 (Biolegend, USA), FITC anti-mouse CD8 (Biolegend, USA), PerCP/Cyanine5.5 anti-mouse CD4 (Biolegend, USA), FITC anti-mouse CD11c (Biolegend, USA), PE anti-mouse CD86 (Biolegend, USA), PE/Cyanine7 anti-mouse CD80 (Biolegend, USA), PE anti-mouse CD44 (Biolegend, USA), PE/Cyanine7 anti-mouse CD62L (Biolegend, USA), PerCP/Cyanine5.5 anti-mouse PD-1 (Biolegend, USA), PE/Cyanine7 anti-mouse OX40 (Biolegend, USA), FITC anti-mouse CD11b (Biolegend, USA), PerCP/Cyanine5.5 anti-mouse F4/80 (Biolegend, USA), APC anti-mouse CD206 (Biolegend, USA), Mouse Regulatory T cell staining kit (eBioscience, USA).

Tumors were enzymatically digested to single cell suspensions and filtered twice through 70 μm filters. Cells were then resuspended in staining buffer (DPBS containing 3% FBS) and stained with the indicated antibodies: APC/Cyanine7 anti-mouse CD45 (Cat. 147717, BioLegend), PE anti-mouse CD3 (Cat. 100205, BioLegend), APC anti-mouse CD8a (Cat. 100711, BioLegend), PerCP/Cyanine5.5 anti-human/mouse Granzyme B (Cat. 372211, BioLegend), VioletFluor™ 450 Anti-Mouse IFN gamma (Cat 75-7311-U025, Tonbo Bioscience), Brilliant Violet 510™ anti-mouse/human CD11b (Cat. 101245, BioLegend), PE anti-mouse Ly-6C (Cat. 128007, BioLegend), FITC anti-mouse Ly-6G (Cat.127605, BioLegend), PerCP/Cyanine5.5 anti-mouse F4/80 (Cat. 123127, BioLegend). Flow cytometry was performed on BD FACS Symphony machines. Flow analyses were performed using FlowJo 10.6.1 software (TreeStar). All flow antibodies were used at a 1:200 dilution.