FIG 3 .

Distinct single-point mutations in 3A render EMCV less dependent on PI4KA. (A to C) Growth curve analysis of EMCV wt and mutant strains. After infection at MOI 1 for 30 min, the inoculum was removed and fresh medium was added to the cells. At the indicated time points, cells were freeze-thawed to determine the total virus titers by endpoint dilution. (A) Growth curves in the absence of compounds in HeLa R19 cells. (B) Growth curves in Huh7-Lunet/T7-shNT and Huh7-Lunet/T7-shPI4KA cells. (C) Growth curves in HeLa R19 cells in the presence of DMSO, 10 µM AL-9, or dipyridamole (Dip). (D) RNA replication of EMCV mutants is resistant to PI4KA inhibitors. HeLa R19 cells were infected at MOI 1 with wt or mutant EMCV reporter viruses carrying the Renilla luciferase gene. After 30 min, virus was removed and compound-containing medium was added to the cells. At 8 h p.i., cells were lysed to determine the intracellular luciferase activity. The value for DMSO samples was set at 100%. Mean values ± SEM are shown. Means were statistically compared using unpaired t tests. **, P < 0.01; ***, P < 0.001.