Fig. 3 | Nature Communications

Fig. 3

From: RSV glycoprotein and genomic RNA dynamics reveal filament assembly prior to the plasma membrane

Fig. 3

SBA allows for the visualization of RSV G dynamics in RSV infected cells. a Vero cells were infected with RSV A2 at an MOI of 1. At 12 hpi, cells were stained live with SBA-488 (top) or vehicle control (bottom), washed, and further incubated in complete growth media. 24 hpi, cells were fixed and immunostained for RSV N (blue), RSV F (red), and nuclei (gray). Scale bar represents 10 µm. b Enlarged cropped images from the white boxed regions from part a with intensity profiles showing colocalization along the direction of the white arrow. c Vero cells were infected with RSV A2 at an MOI of 1. 12 hpi, cells were stained live with SBA-488 (top, green) or vehicle control (bottom), washed, and further incubated in complete growth media. 24 hpi, virus was harvested. Filaments were isolated onto glass and stained for RSV F (blue) and RSV G (red) Scale bar represents 2 µm. d Mean sum of RSV G intensity per filament was quantified. 50 isolated virions were analyzed for each condition. Error bars represent standard deviation. There was no statistically significant difference between SBA-labeled and unlabeled RSV (Mann–Whitney U test, p > 0.05). e Vero cells were infected with RSV A2 at an MOI of 1. 12 hpi, cells were stained live with SBA-488 or vehicle control, washed, and further incubated in complete growth media. 24 hpi, supernatants from single infected wells were titered by plaque assay in quadruplicate. Three wells were independently infected, stained, and assayed per treatment. Error bars represent standard deviation. There was no statistically significant difference between SBA-labeled and unlabeled RSV (Mann–Whitney U test, p > 0.05)