FIG 6.

A cholesterol efflux block impairs viral exit from endosomes. (A) Top panels, Rab7-labeled endosomes in control untreated, uninfected cells (DMSO), aggregates of dilated endosomes in uninfected cells treated with U18666A, and control infected, untreated cells. Virions are labeled with anti-p72 antibody, and endosomes labeled with Rab7 marker are shown in green. Bottom panels, U18666A-pretreated cells infected with ASFV at 1 hpi. Dilated perinuclear endosomes filled with cholesterol shown in black and white in an independent channel. Arrowheads and zoom images show viral capsid staining of intact virions retained at the periphery of dilated endosomes. Scale bars, 10 μm. (B) Number of virions found inside endosomes in control and U18666A-treated cells (***, P < 0.001). (C) Effect of cholesterol flux inhibition on ASFV, VV, and Adv infectivity evaluated by flow cytometry (***, P < 0.001; *, P < 0.05).