PANC-1, HepG2, SGC-7901, and MDA-MB-231 cells infected with sh-Control or sh-ITGA2s were prepared and stained for flow cytometry with the following antibodies: isotype APC anti-human IgG Fc Antibody (Biolegend, clone HP6017, USA) or APC anti-human CD274 antibody (Biolegend, clone 29E.2A3, USA) or for 30 mins at 4 °C. The cells were then washed with PBS three times, resuspended in 150 μl staining buffer, and analyzed for flow cytometry.
For flow cytometry analysis of the mouse tissue samples, single-cell suspensions were prepared and stained for flow cytometry with the following antibodies: APC conjugated CD45 antibody (Biolegend, 103,112, USA); FITC conjugated CD4 antibody (Biolegend, 100,510, USA); PE-conjugated CD8 antibody (Biolegend, 100,708, USA); APC conjugated CD11b antibody (Biolegend,101,212, USA); and FITC conjugated Gr1 antibody (Biolegend, 108,406, USA). Flow cytometry was performed on BD FACSCelesta (BD Biosciences, USA), and the data were analyzed using FlowJo.
For flow cytometry analysis of the mouse tissue samples, single-cell suspensions were prepared and stained for flow cytometry with the following antibodies: APC conjugated CD45 antibody (Biolegend, 103,112, USA); FITC conjugated CD4 antibody (Biolegend, 100,510, USA); PE-conjugated CD8 antibody (Biolegend, 100,708, USA); APC conjugated CD11b antibody (Biolegend,101,212, USA); and FITC conjugated Gr1 antibody (Biolegend, 108,406, USA). Flow cytometry was performed on BD FACSCelesta (BD Biosciences, USA), and the data were analyzed using FlowJo.