Modulation of the Host Lipid Landscape to Promote RNA Virus Replication: The Picornavirus Encephalomyocarditis Virus Converges on the Pathway Used by Hepatitis C Virus (original) (raw)
Fig 5
PI4P homeostasis is affected upon EMCV infection.
(A) EMCV alters the distribution of intracellular PI4P lipids. Huh7-Lunet/T7 cells were mock-infected or infected with GFP-EMCV at an MOI of 10. At 6 h p.i., cells were fixed and stained with antibodies against PI4P using specific protocols for detection of plasma membrane and intracellular PI4P pools, as described previously [50]. Nuclei were stained with DAPI (blue). (B) Quantification of PI4P levels by immunofluorescence analysis. Huh7-Lunet/T7 cells were mock-infected or infected with EMCV at an MOI of 10. At 6 h p.i., cells were fixed and stained for intracellular PI4P and viral dsRNA. The intensities of fluorescent PI4P signals from whole-cell z-stacks were quantified using ImageJ. Shown are mean values ± SEM of 30 cells per condition. Means of mock and infected cells were statistically analyzed using the Mann–Whitney test. ***P<0.001. (C) EMCV ROs contain PI4P. Huh7-Lunet/T7 cells were mock-infected or infected with EMCV-2C-HA at an MOI of 10. At 5.5 h p.i., cells were treated with DMSO or 1 μM BF738735 (PI4KB inhibitor) for 30 min, then fixed and stained for intracellular PI4P and HA using specific antibodies. Nuclei were stained with DAPI (blue). The crop panels at the bottom depict an enlargement of the boxed areas. Scale bars represent 10 μm.