The Ubiquitin/Proteasome System Mediates Entry and Endosomal Trafficking of Kaposi's Sarcoma-Associated Herpesvirus in Endothelial Cells
Figure 5
Inhibition of proteasome function reduces KSHV particles associated with late endosomal/lysosomal marker LAMP1.
(A) HUVEC treated with DMSO, MG132 or EPOX were incubated with KSHV for 4 hr, stained for KSHV particles (red), LAMP1 (green), membrane (white), and nuclei (blue). Z-stacks were deconvolved and, Imaris image analysis software was used to generate 3-D contoured images. Videos S9, S10 and S11 correspond to cells treated with DMSO, MG132 or EPOX, respectively, labeled for KSHV particles and LAMP1 rotated around the x-axis. Video S12 depicts KSHV particles enclosed in an LAMP1+ vesicle. (B–D) Imaris 3-D colocalization analyses determine the total numbers of KSHV particles in a cell that are colocalized with LAMP1 (B), the numbers of viral particles localized at cell membranes that are LAMP1+ (C), and the numbers of viral particles localized within the cytoplasmic spaces that are LAMP1+ (D). Box and whisker plots depict the statistical analyses of the cellular localization of KSHV particles as described in Figure 3.