Fig 8. Subcellular localization of proteins biotinylated by BioID-L.

(A) Confocal microscopy images of biotinylated proteins stained with streptavidin-Alexa Fluor 594 (orange) and 3D viral polymerase (green) in HeLa cells infected for 10h with BioID-L^WT, BioID-L^M60V or BioID-L^F48A replicons (MOI 5). 3D viral polymerase was stained as a control of infection. (B) Quantification of nuclear rim staining in HeLa cells infected with BioID-L^WT, BioID-L^M60V or BioID- L^F48A viruses (mean ± SD). Counts: 31 ± 11 infected cells per experiment (n = 4). One-way ANOVA was used to compare all samples with each other. Significant differences are shown. (C) BioID-L biotinylates proteins that colocalize with the FG-NUP POM121. Confocal microscopy images and intensity vs distance plots of biotinylated proteins (orange) and POM121 (green) in HeLa cells infected with BioID-L^WT, BioID-L^M60V or BioID-L^F48A viruses for 10h (MOI 5). White arrows point to examples of cells exhibiting a nuclear rim staining. Scale bar: 10μm.