Fig. 2: Characterization of the azido-labeled A375 EVs.
From: Capturing nascent extracellular vesicles by metabolic glycan labeling-assisted microfluidics
a Schematic diagram of the click reaction between the azido-modified A375 EVs and the alkyl dye DBCO-Cy5. b Fluorescence intensity of the A375 EVs treated with different concentrations of Ac4ManNAz (left) or DBCO-Cy5 (right). The EVs were loaded on latex beads, and then reacted with the DBCO-Cy5 dye prior to characterization. c Morphology comparison of the MGL A375 EVs and non-MGL A375 EVs by TEM. Scale bar = 200 nm. d, e Comparison of the size distribution (d) and Zeta potential (e) of the MGL (w/ MGL) A375 EVs and non-MGL (w/o MGL) A375 EVs by dynamic light scattering. n = 3 biologically independent experiments. Data shown as mean ± SD. f Characterization of CD63 expression on the azido-labeled A375 EVs. g Veen diagram to compare the A375 EV proteins before and after MGL. h–j Biological processes (h), cellular components (i) and molecular functions (j) of gene ontology (GO) enrichment analysis of proteins detected in A375 EVs with (in orange) or without (in gray) MGL. Proteins co-expressed in both are indicated in light orange. k Heatmap of the expression levels of the 806 proteins detected both in azido-labeled or unlabeled A375 EVs. The log 10 expression values for the overlapping proteins are indicated by the colors shown in the scale. l Heatmap with the top 30 proteins (in triplicate) of EVs from A375 cells with (in orange) or without (in gray) Ac4ManNAz treatment. The expression changes are indicated in log 2 scale. Source data are provided as a Source Data file.
