Figure 7

PSaV entry is pH-dependent and involves actin and microtubules. A, B Mock- or chloroquine-treated LLC-PK cells were inoculated with the PSaV Cowden strain and then incubated with CMFDA to visualize acidification of intracellular compartments, and incubated for further 30 min in maintenance media (A) or incubated with CMFDA followed by LAMP2 antibody to check colocalization by confocal microscopy (B). C, D After pretreatment of LLC-PK cells with chloroquine and infection with AF594-labeled PSaV particles (approximately 415 particles per cell), cells were incubated with neutral (pH 7.2) (C) or acidic (pH 5.2) buffers (D). The cells were prepared for confocal microscopy to check colocalization of AF594-labeled PSaV particles with LAMP2. All experiments were performed in triplicate and panels A–D show representative sets of results. Scale bars for panels A–D correspond to 10 μm.