Fig. 1

Characterization of exosomes derived from microglia treated with or without ethanol. Nanoparticle tracking analysis (A, B), Mean ± SEM data of exosome concentration (C, D), Transmission electron microscopy (E), and Western blot verification (F) of exosomes prepared from media of hypothalamic microglial cultures treated with vehicle (Control) or 50 mM ethanol (Ethanol) for 24 h, or (G) from microglia of mediobasal hypothalamus (MBH) tissue obtained from PND 6 male or female rats, which were fed daily from PND 2–6 with ethanol-containing milk formula (AF), pair-fed isocaloric milk formula (PF), or left undisturbed in the litter with mother (AD). For nanoparticle tracking analysis, the calculated size distribution is depicted as a mean (black line) with SE (red shaded area). Mean particle size, mode particle size, and concentration of particles in exosomes are shown in each tracking analysis graphs. Ethanol -induced changes in exosome concentration in primary microglia in culture and MBH microglia in tissue preparation are shown in panel C and D, respectively.Representative images of exosomes from MBH microglia of AD rats under TEM (E). The width and length of each exosome are represented on the individual exosome. Representative bands for the protein levels of exosome proteins TSG101 and Alix, mitochondria marker cytochrome c, Golgi apparatus marker GM130, and endoplasmic reticulum marker calnexin in exosomes from primary microglia and MBH microglia as determined by Western blot analysis (F, G). Data shown in Fig. 1C are mean ± SEM (N = 4) and were analyzed with unpaired t-test while the data shown in Fig. 1D are mean ± SEM (N = 3) and were analyzed using two-way analysis of variance with Tukey’s multiple comparison post hoc test. *, p < 0.05; **, p < 0.01