Inhibition of proteasome function reduces KSHV particles associated with late endosomal/lysosomal marker LAMP1.

<p>(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. <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1002703#ppat.1002703.s013" target="_blank">Videos S9</a>, <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1002703#ppat.1002703.s014" target="_blank">S10</a> and <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1002703#ppat.1002703.s015" target="_blank">S11</a> correspond to cells treated with DMSO, MG132 or EPOX, respectively, labeled for KSHV particles and LAMP1 rotated around the x-axis. <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1002703#ppat.1002703.s016" target="_blank">Video S12</a> 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 <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1002703#ppat-1002703-g003" target="_blank">Figure 3</a>.