Figure 3.

PI4KIIIβ Is Localized to Enteroviral RNA Replication Organelles

(A) PI4KIIIβ and Arf1 are colocalized throughout CVB3 infection. HeLa cells expressing Arf1-GFP infected with CVB3 were immunostained with anti-PI4KIIIβ and anti-GFP antibodies.

(B) GBF1/Arf1 inactivation leads to PI4KIIIβ dispersal. HeLa cells were infected with CVB3 for 4 hr and treated with 10 μg/ml of BFA for 30 min. 75% ± 5% (n = 10 cells) of PI4KIIIβ-associated fluorescence was dispersed upon BFA treatment.

(C) PI4KIIIβ is in physical complex with viral replication enzymes. HeLa cell lysates from cells infected with CVB3 or mock (−) for 4 hr were immunoprecipitated with anti-PI4KIIIβ antibodies. Input samples verifying the presence of TGN46 in the lysates prior to immunoprecipitation are presented in the bottom panel.

All fluorescence images were confocal images of optical slice thickness ∼1 μm. Scale bar, 10 μm.