Figure 5.

Internalisation of schistosomula EVs without surface N-glycans via DC-SIGN and EV-associated glycolipid-glycans.

The geoMFI (geometric mean fluorescent intensity) of PKH26 relative to unstimulated moDC after 2 h incubation with labelled EVs that were treated without or with PNGase F (a). Pre-incubations are indicated on the x-axis. MoDC from 3 donors. Mean±SEM *p < 0.05, ***p < 0.001, using repeated measures ANOVA with Tukey’s Multiple Comparison Test. EV-derived glycolipid-glycans measured by MALDI-TOF-MS (b). Signals are labelled with monoisotopic masses. Putative structural assignments were deduced from these masses based on published data of schistosomula glycans [34] and on hydrofluoric acid (HF) data (not shown). Coloured bars indicate the corresponding core structure with varying numbers of fucose substitutions. Putative terminal motifs formed by these fucoses are indicated in the insert. The spectrum shown is representative for three biological replicates. Red triangle, fucose; yellow circle, galactose; blue square, N-acetylglucosamine; yellow square, N-acetylgalactosamine; LeX, LewisX; LeY, LewisY; F-LDN/LDN-F/F-LDN-F, fucosylated LacDiNAc *, signals corresponding to a hexose oligomer of unknown origin; #, non-glycan signals