Anti apccy7 cd45

The collected tumor tissues were prepared as single cell suspensions, stained with Zombie Aqua Fixable Viability Kit (anti-BV510, BioLegend, #423102, USA), incubated for 15 min at room temperature, and then terminated and centrifuged. The fragment crystallizable (FC) was blocked with anti-CD16/32 (BioLegend, #101320) and stained with surface antibody after 15 min (in two tubes, the first tube was stained as follows: anti-PE-NK1.1 (BioLegend, #108708); anti-PC5.5-CD4 (BioLegend, #100434); anti PECY7-CD8 (BioLegend, #100722); anti-APC-CD3 (BioLegend, #100236); anti-APCCY7-CD45 (BioLegend, #103116); anti-BV711-interferon (IFN)-γ (BioLegend, #505836). Tube 2: anti-fluoresceine isothiocyanate (FITC)-major histocompatibility complex (MHC)-II (BioLegend, #107606); anti-PE-CD11B (BioLegend, #101208); anti-PC5.5-Gr1; anti-APC-F4/80 (BioLegend, #123116); anti-APCCY7-CD45), incubated at 4°C for 30 min, followed by fixed membrane breaking solution incubation for 30 min, and stained with intracellular antibodies (tube 1: anti-FITC-granzyme B (BioLegend, #372206); anti-BV711-IFN-γ, tube 2: anti-PECY7-CD206 (BioLegend, #372206)) incubated at 4°C for 30 min, followed by flow cytometry detection (BD FACS LSRFortessa, USA).