Fig. 5

Viral inclusions are associated with ER exit sites. a Sec61β-Emerald cells were transfected with mCherry-NP and infected with PR8 virus, at an MOI of 10, for 16 h. b A549 cells were co-transfected with plasmids encoding mCherry-NP and ER-GFP and infected with PR8 virus, at an MOI of 10, for 16 h. a, b Cells were imaged under time-lapse conditions. Individual frames with single moving particles highlighted with yellow arrows are shown. Images were extracted from Supplementary Movies 17 and 18. More than 15 cells from 3 independent experiments were analyzed in each condition. c A549 cells were infected or mock-infected (M) with PR8 virus, at an MOI of 3, and fixed at the indicated times. Cells were stained for the ER proteins Sec31A (in green) and PDI (in gray) and the viral NP protein (in red). Areas highlighted by the white box are shown on the right of each panel. Yellow arrowheads indicate co-localization between Sec31A and NP. Bar = 10 μm. More than 30 cells from 2 independent experiments were analyzed. d A549 cells were co-transfected with plasmids encoding mCherry-NP and GFP-Sec16 and infected or mock-infected (M) with PR8 virus for 16 h. Cells were imaged under time-lapse conditions. Representative cells are shown in the left large images. Individual frames with single moving particles highlighted with yellow arrows are shown in the small panels. Two examples are provided for the infected cell (16 h.1 and 16 h.2). Bar = 7.5 μm. Images were extracted from Supplementary Movies 19 and 20. Images are representative of at least 15 cells, from 2 independent experiments