FIG 6 .

EMCV mutants do not accumulate OSBP and cholesterol at the ROs in the presence of PI4KA inhibitors. HeLa R19 cells were infected with EMCV-3A-A32V for 30 min at MOI 10, followed by treatment with PI4KA inhibitor AL-9 where indicated. At 6 h p.i., cells were fixed and stained for 3AB, OSBP, and cholesterol. Cholesterol was visualized using filipin, a fluorescent antibiotic that specifically binds free cholesterol. For clarity, the overlay panels depict the outline of the cell (white line). Colocalization of OSBP or cholesterol with 3AB was determined by calculating the Pearson’s correlation coefficients for at least 15 cells for each condition. Mean values ± SEM are shown. Means were statistically compared using the Mann-Whitney test. ***, P < 0.001. Scale bars represent 10 µm.