Fig. 5 .

Microglial activation by sepsis plasma EVs induces neuronal apoptosis. A Schematic illustration of experimental setup. Conditioned media (CM) were collected at 24 h from microglia treated with plasma CLP- or sham-EVs using aseptic technique. The primarily cultured neurons were incubated with either sham or CLP EVs directly or CM collected above and apoptosis were evaluated at 16 h using western blot and fluoresce imaging. B Conditioned media from CLP EVs treated-microglia but not CLP EVs itself induce apoptosis to neurons. Representative images of blot from three different experiments using different batches of primary neuronal cultures. C Relative intensity of cleaved caspase-3 over caspase-3 precursor. n = 4–10/group D Annexin V-staining in neurons. Neurons were incubated with the conditioned media from EV-treated microglia for 16 h and Annexin V-FITC was stained. Representative images from three different experiments