Fig. 1
From: Extracellular vesicles during Herpes Simplex Virus type 1 infection: an inquire
Separation of STING-carrying vesicles from HSV-1 virions. (a) Supernatant from human epithelial cells (HEp-2) infected with HSV-1(F) was clarified by differential centrifugation to remove cell debris and nuclei, filtered concentrated before loaded onto a iodixanol gradient, as detailed in Deschamps T. and Kalamvoki M, manuscript in preparation. Fractions were collected from the top to the bottom of the gradient and the proteins were identified by immunoblot analysis. The tegument virion protein 22 (VP22) and the capsid unique long 38 protein (UL38) were found in high density iodixanol fractions. STING, CD63 and CD9 were floating in the low density fractions. (b) The same fractions were tested for the presence of infectious viral particles, by plaque assay in Vero cells. The number of viral plaques in each fraction were counted after Giemsa staining. (c) EVs and virions derived from the supernatant of HEp-2 cells exposed to HSV-1(F) were pelleted before loaded onto a dextran-10 gradient (1.04-1.09Â g/cm3). The HSV-1 virions and the tetraspanin CD9 along with STING were found in the same fraction