FIG. 3.

KSHV trafficking after treatment with macropinocytic inhibitor EIPA. HMVEC-d cells grown in eight-chamber slides were left untreated or treated with 0.25 μg EIPA, washed, infected with KSHV at an MOI of 100 for 30 min (30′) at room temperature, washed, fixed, permeabilized, blocked in 5% BSA, incubated in BSA with primary antibodies to tubulin and KSHV capsid ORF65 protein for 1 h at room temperature, washed, incubated with secondary antibodies to tubulin (green) and ORF65 (red), washed, mounted, and viewed under an immunofluorescence microscope. Pink arrowheads indicate KSHV capsids, red arrows indicate tubulin, and white arrowheads indicate colocalization of KSHV capsids with microtubules. Magnification, ×80. Boxed areas in c and g are shown enlarged in d and h.