Figure 5

Confocal and super-resolution microscopy: PaKi cells were infected with HeV, fixed at 18 hpi and prepared as for conventional confocal imaging. a, b) Cells were labelled with anti- HeV G antibody detected with anti-human antibodies conjugated to Alexa 568 (a, red) and co-localised with rabbit anti-N antibody detected with anti-rabbit antibodies conjugated to Alexa 488 (b, green). The labelling is co-localised in dots and there is also N protein labelling within the cell cytoplasm (arrow). Cell structure is shown in image b) using DIC. c) SR GSD image of a similar culture labelled as for a, b. HeV G protein (detected with Alexa 647; (shown as green for consistency) can be seen as round circles around the viral particles and HeV N protein (Alexa 488, shown as red for consistency) is inside the particles. d) SIM image of cells prepared as for a, b). The G protein (green) forms a ring around the particles with the N protein (red) inside. The arrow indicates an area of G protein labelling on the cell membrane. Scale bars: a, b) =5 μm, c, d) =500 nm.