Fig. 2 | Nature Communications

Fig. 2

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

Fig. 2

SBA binding does not inhibit RSV binding or replication. a SBA-488 (green) was incubated with RSV A2 stocks for 30 min before filaments were isolated onto glass coverslips. Isolated virions were fixed and stained for RSV N (red) and RSV F (blue). Scale bar represents 2 µm. b SBA-488 labeled virus (green) was used to inoculate Vero cells at an MOI of 1. At 24 hpi, cells were fixed and immunostained for RSV N (red), RSV F (blue), and nuclei (gray). Scale bar represents 10 µm. c RSV A2 was incubated with either SBA or vehicle control for 2 h at 37 °C. Virus was then used to inoculate Vero cells at an MOI of 1. At 12 or 24 hpi, cells were lysed, and lysates were assayed by western blotting for RSV G. β-actin is included as a loading control. Image is representative of two independent experiments. d Replicate lanes (n = 2) were quantified by densitometry to compare the amount of RSV G signal from SBA-labeled or unlabeled RSV infections. There were no statistically significant differences at either time point (two-way ANOVA, p > 0.05). Error bars represent standard deviation. e rgRSV-GFP was incubated with the indicated concentrations of either palivizumab, SBA, or vehicle control for 2 h at 37 °C. Virus treatments and a mock treatment were then used to inoculate Vero cells at an MOI of 1. At 48 hpi cells were lifted into a single-cell suspension and analyzed by flow cytometry. Data represents the mean percentage of infected cells in three independent experiments. Error bars represent standard deviation. Asterisk indicates significant difference vs. mock infection (one-way ANOVA on ranks, p < 0.05). f Relative quantity of RSV N (left) and RSV G (right) expression via qRT-PCR upon infection of Vero cells with SBA-labeled or unlabeled RSV A2 at an MOI of 1 at 6, 12, and 24 hpi. There was no statistically significant difference at any time point (two-way ANOVA, p > 0.05). Error bars represent standard deviation. Results are the mean of two independent experiments normalized by the control virus at 24 hpi