Figure 6.

Glycan modification of EVs improved internalization by DCs. (a). Dot plots showing binding/uptake of unmodified or Le^Y-incorporated DiD⁺ EVs by moDCs after 30 min incubation at 4°C (T = 0) or after 90 min incubation at 37°C (T = 90) measured by FACS, representative of three donors. (b). Percentage DiD⁺ moDCs after incubation with unmodified or Le^Y-incorporated DiD⁺ EVs for 30 min at 4°C (T = 0). MoDCs were fixed after 0, 30, 60, or 90 min incubation at 37°C and DiD signal was measured by imaging flow cytometry (three donors). (c). Binding/uptake of EVs by moDCs (three donors), measured by imaging flow cytometry. (d). Representative imaging flow cytometry brightfield (BF) pictures of moDCs with DiD positive spots on the outside and inside of cells. Blue area masks inside area of cells, and colocalization of DiD signal with the mask was used to calculate internalization scores. (e). Internalization of EVs by moDCs in a pulse-chase experiment with cells fixed in PFA after 15, 30, 45 and 60 min incubation at 37°C visualized in one donor by imaging flow cytometry.