Apc anti human cd19 clone hib19

After incubation with EVs, the PBMCs were washed three times in FACS buffer (0.5% BSA in PBS) and then stained with fluorescent labeled monoclonal antibodies from Biolegend: APC anti-human CD19 (clone HIB19), FITC anti-mouse/human CD11b (clone M1/70), FITC anti-human CD14 (clone M5E2), PerCP/Cy5.5 anti-human CD25 Antibody (clone BC96), PE/Cy7 anti-human HLA-DR Antibody (clone L243), and Pacific Blue™ anti-human CD3 (clone HIT3a) or PerCP/Cy5.5 anti-CD4 (clone OKT4), and PE/cy7 anti-human CD8 (clone RPA-T8), according to manufacturer’s recommendations. In Fig. 4, we used additional antibodies including BV422 anti-human CD80 (clone 2D10), Alexa Fluor 700 CD15 (cloneW6D3), FITC anti-human CD14 (M5E2), and AmCyan CD11b (clone M1/70). After antibody incubation, cells were washed two times in FACS buffer. Flow cytometry data was collected on a FACS Canto II (BD, Franklin Lakes, NJ). Dead cells were excluded from analysis using forward and sideward scattering gating. Live cells were gated based on cell surface markers CD11b, CD14, CD19, CD4 and CD8 or CD3 to identify monocytes, B cells, and T cells, respectively. For Fig. 4, percent monocytes are shown as CD14⁺CD15⁻ cells gated on CD11b⁺ population. The cell populations were then also gated for PKH to identify cells that internalized the labeled EVs. The flow cytometry data was analyzed with FlowJo software (Tree Star, Inc.).