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. 2015 Jan 8;11(1):e1004597. doi: 10.1371/journal.ppat.1004597

Figure 11. KSHV-mediated p-body dispersion in latently infected endothelial cells requires kaposin expression.

Figure 11

A, D) HUVEC cells were either not infected or infected with KSHV for 24, 48, or 72 hours before being fixed with 4% paraformaldehyde. Cells were permeabilized in 0.1% Triton X-100 (in PBS), and blocked in 1% human AB serum before staining with the following primary antibodies: anti-hedls (p-bodies, green) and anti-LANA (to mark infected cells, red). In part A, infected cells are denoted with a thin arrow; uninfected cells are marked with a thick arrow. In part D, actin stress fibers were also labeled with phalloidin (false-colored white). B–C) To quantify the effect of latent infection on p-bodies, the number of LANA-expressing cells that displayed p-bodies of normal size (>300 nm in diameter) were counted and compared to uninfected cells. Three independent experiments were performed. >100 cells were counted in each experiment. C–D) Before KSHV infection HUVECs were transduced with two different recombinant lentiviruses that express a short hairpin RNAs (shRNAs) against the kaposin transcript (named shKap1 and shKap2) or the non-specific (NS) shRNA control. Positive transductants were selected by puromycin treatment for two days and then seeded on coverslips for infection with KSHV the next day. 48 hours post-infection, cells were fixed and processed for immune fluorescence as described above. n = 3 independent experiments Scale bar  = 10 µm.