FIG 4 .

EMCV mutants recruit PI4KA to their ROs. (A) HeLa R19 cells were transfected with a plasmid encoding GFP-PI4KA. The next day, cells were mock infected or infected with CVB3 (included as negative control) or the EMCV wt or mutants at high MOI. At 6 h postinfection (p.i.), cells were fixed and stained with antibodies against EMCV 3AB or CVB3 3A as RO markers. (B) 3A-A32V and 3A-A34V alone can recruit PI4KA to membranes. Huh7-Lunet/T7 cells were cotransfected with the GFP-PI4KA construct and either empty vector or plasmids encoding myc-tagged 3A of CVB3 (included as negative control) or 3A of wt or mutant EMCV. The next day, cells were fixed and stained with antibodies against the myc tag to detect the overexpressed 3A proteins. Insets depict enlargements of boxed areas. (A and B) Nuclei were stained with DAPI (blue). Scale bars represent 10 µm.