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. 2021 Jul 8;12:627394. doi: 10.3389/fphar.2021.627394

FIGURE 4.

FIGURE 4

The vimentin-binding compound R491 inhibited exosomes secretion rather than exosomes generation. (A) Representative transmission electron micrographs of exosomes (red arrows) purified from A549 and PANC-1 cells treated with DMSO or R491 at 5 µM scale bars: 100 nm. (B) Nanoparticle tracking analysis (NTA) of small extracellular vesicles (sEVs) released from A549 and PANC-1 cells treated with DMSO or R491 at 1 µM sEVs were isolated by serial ultracentrifugation from cell culture supernatants of same number of cells. (C) NTA quantification of three independent experiments. (D) Western blot analysis of A549 cells treated with DMSO or with R491 at 0.1, 1, and 10 µM for 48 h. Extracts from cells and sEVs (Exo) were blotted for the exosomal markers CD9, CD63, and TSG101. (E) Quantification of exosomal proteins extracted from sEVs of A549 cells in three independent experiments. (F) Western blot analysis of PANC-1 cells treated with DMSO or with R491 at 0.1, 1 and 10 µM for 48 h. Extracts from cells and sEVs (Exo) were blotted for the exosomal markers CD9, CD63 and TSG101. (G) Quantification of exosomal proteins extracted from sEVs of PANC-1 cells in three independent experiments.