Figure 1.

Characterization of EVs Isolated from Astrocyte Culture

(A) Schematic representation of the major steps involved in the EV isolation from astrocyte culture. (B) Western blot characterization of astrocyte EVs. Protein isolated from astrocyte EVs was separated on SDS-PAGE and electroblotted onto nitrocellulose membrane. Blots were probed with exosome marker antibody against TSG101, CD63, and Alix. Calnexin was used as a control for cell debris contamination. (C) Representative electron micrograph of EVs isolated from A172 cells. Scale bar, 200 nm. (D) AFM image of EVs isolated from mouse primary astrocyte culture supernatants. (E) Size and particle distribution plots of isolated EVs from cell culture by NanoSight Tracking Analysis (NTA). The plot shows a peak size around 80 nm for the isolated EVs. (F) Number of EVs isolated from control and morphine-exposed human primary astrocytes. All experiments were done at least three independent times. Data are shown as mean ± SEM. *p < 0.05 versus control.