Quantifying Surface CD8-CI-MPR Expression

To assess the levels of surface CD8-CI-MPR, unfixed cells were incubated for 30 min at 4 °C with mouse anti-CD8 antibody (9 (link), 59 (link)) followed by incubation for 30 min at 4 °C with F(ab')2-Goat anti-Mouse IgG (H + L) Cross-Adsorbed Secondary Antibody Alexa Fluor 647 (Cat. A21237; Thermo Scientific) in PBS supplemented with 1% BSA, and then fixed with PFA 1% in PBS + 1% BSA and subjected to flow cytometry analysis. Data were acquired using the levels of Alexa 647 fluorescence in cells from a BD LSRFortessa flow cytometer (BD Biosciences) and FACSymphony A5 Cell Analyzer (BD Biosciences) at the Ribeirão Preto Center for Cell-Based Therapy/Hemotherapy. The FloJo software (Tree Star) was used for data analyses.