Figure 1.

Characterization of ReN cell-derived EVs. (A) Transmission electron micrograph of EV_ReN. Scale bar, 100 nm. (B) Size distributions of EV_ReN based on NTA measurements. (C) Western blot analysis of Alix, TSG101, and Calnexin from ReN cells and EVs isolated from their conditioned medium. The supernatant obtained from the ultracentrifugation during EV isolation was used as a negative control. (D) Representative confocal images of cellular uptake of tdTomato-labeled EV_ReN after 2-h incubation with BV2 microglia. Red shows EVs. Green indicates cell profile. Blue is nuclei. Arrows indicate the EVs in cytoplasma. Scale bar, 30 µm. (E, F) LPS-stimulation was performed on BV2 microglia receiving 24 h of PBS, EV₂₉₃, or EV_ReN treatments. (E) RT-PCR analyses (n = 4) of TNF-α, IL-1β, and IL-6 in the cells were performed 1 h after LPS-induction. (F) ELISA analyses (n = 5) of TNF-α, IL-1β, and IL-6 in the conditioned medium were performed 24 h after LPS-induction. Data are expressed as mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001 versus the LPS-stimulation without treatment groups by One-way ANOVA.