Fig. 1

Characterization of sEVs released from glutamate or GABA-treated neurons. A, B Primary cultured neurons on DIV 15 were treated with a gradient concentration of glutamate (A) or GABA (B) and subjected to a CCK-8 cell viability assay. C–H Neurons were cultured in a T75 culture flask. The number of cells and the volume of culture medium in the three groups were consistent. After neurons on DIV 15 were treated with 10 μM glutamate (diluted in 20 μL PBS), 300 μM GABA (diluted in 20 μL PBS), or PBS (20 μL) for 24 h, sEVs were extracted by serial ultracentrifugation from cell culture supernatants from equivalent numbers of cells. C The positive transmembrane sEV marker CD63, the cytosolic sEV marker Tsg101, and the negative controls GM130, COX IV and TOMM20 were detected in the sEVs and their derived cells by Western blot. sEVs: Neuron-derived sEVs, TCL: Total cell lysates. D Quantification and statistics of CD63 and Tsg101 protein levels detected by Western blot from three independent experiments. E Quantification and statistics of the concentration of the nanoparticles detected by nanoparticle tracking analysis (NTA) from three independent experiments. F Representative NTA traces of sEVs were derived from three experimental groups. G The total protein amount of the sEVs in each group were detected by the BCA method. H The morphology of sEVs from PBS-treated neurons was observed under a transmission electron microscope. n = 3. Data are presented as the mean ± SEM, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001