Fig. 1: LPS treatment had no effect on EVs release in CECs but increased microglial uptake of CEC-EVs. | Cell Death & Disease

Fig. 1: LPS treatment had no effect on EVs release in CECs but increased microglial uptake of CEC-EVs.

From: Cerebral endothelial cell-derived extracellular vesicles regulate microglial polarization and promote autophagy via delivery of miR-672-5p

Fig. 1

A Schematic diagram of EVs isolation from conditioned media. B Size distribution of EVs in 2 K, 10 K and 100 K pellets. C Total protein concentrations of 2 K, 10 K and 100 K pellets derived from control group and LPS group. D 2 K, 10 K and 100 K pellets were blotted for the exosomal markers CD63 and syntenin, and for the endoplasmic reticulum marker GP96 and Calnexin. E Flow charts for the EVs purification procedure based on sucrose density gradient centrifugation. F Western blot for exosomal markers CD63, syntenin and Calnexin in different sucrose gradient fractions. G Representative imaging distribution of the DiR-labeled CEC-EVs in different organs, including brain, heart, liver, spleen, lung, and kidney. H CEC-EVs can cross the blood-brain barrier and neuroinflammatory markedly enhanced the incorporation of CEC-EVs into microglia. I Transwell co-culture system. J LPS treatment increased microglial uptake of CEC-EVs. The lipid membrane of CECs was stained with PKH26 red fluorescent dye and the cells were co-cultured with BV2 microglial cells stained with DIO in a Transwell. All data are present as means ± SD (n = 3). Unpaired Student’s t- test. ns not significant.

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