Essential Domains of Oxysterol-Binding Protein Required for Poliovirus Replication

Figures

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  Figure 1. OSBP variants analyzed in this study. (Upper) A crystal structure of ligand-binding domain of human OSBP (PDB: 7V62). The M446 residue and a C-terminal region (aa 754–807) of the OSBP is highlighted in magenta and purple, respectively. (Lower) Domain organizations of OSBP variants analyzed in this study. The corresponding amino acid numbers of the domains are shown. IDR: intrinsically disordered region; PHD: pleckstrin homology domain; CCR: coiled-coil region; FFAT: two phenylalanines in an acidic tract.

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  Figure 2. Subcellular localization and expression levels of the OSBP variants. (Left) Maximum projection view of HEK293 cells expressing the OSBP variants analyzed by confocal microscopy. The cells were infected with PV1(mCherry)pv at an MOI of 50 at 37 °C for 5 h. Images of the representative cells (mock-infected or PV1(mCherry)pv-infected cells) at 5 h p.i. are shown. (Right) Expression levels of the OSBP variants. The cells were analyzed by flow cytometry. The expression level of OSBP-EGFP(WT)-expressing cells is taken as 100%. The data are representative of two independent experiments with two or three biological replicates.

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  Figure 3. Functional dominance assay of OSBP. HEK293 cells expressing each of the OSBP variants were infected with PV1(Fluc)pv at an MOI of 0.5 at 37 °C for 7 h in the presence (20 μM) or absence of T-00127-HEV2. PV1(Fluc)pv infection in the absence of T-00127-HEV2 is taken as 100%. Statistical significance of the infection to that in the WT-expressing cells was shown. n.s., not significant. ***, p < 0.001. The data represent the mean and standard deviation of two independent experiments with two or three biological replicates.