Extended Data Fig. 5: RSV IBs lose velocity and the ability to fuse but remain sensitive to 1,6-hexanediol.

a, b, HEp-2 cells infected with RSV–P–BFP for 24 h were treated for 1 h with DMSO, CPM (5 μM) or A3E (25 μM) and then exposed to 10% 1,6-hexanediol (Hex), imaged at 10 min and quantified (see Methods). Data are from 10 acquisitions captured in two independent experiments. Representative images are shown in a. Scale bar, 10 μm. The mean ± s.d. number of IBs per image is expressed as the percentage of the pre-treatment control. **P < 0.01; two-tailed Wilcoxon signed-rank test. c, d, HEp-2 cells were infected with RSV–P–BFP for 24 h and treated for 1 h with DMSO, CPM (5 μM) or A3E (25 μM) and then imaged as described in the Methods. Fusion events were quantified as described in the Methods. Results are expressed as mean ± s.d. from 10 videos (15 min) from two independent experiments (c). Maximum velocities were obtained by automatic tracking of IBs from 10 videos (15 min) from two independent experiments (see Methods). Box plots show the median (centre line) and the first and third quartiles (upper and lower hinges). Statistical analysis of maximum velocities was done using Kruskal–Wallis tests followed by Dunn’s test for multiple comparisons (d). *P < 0.05; ***P < 0.001. e, HEp-2 cells were infected for 24 h with RSV–P–BFP and treated with CPM (5 μM), A3E (25 μM) or DMSO as control for 1 h. IBs shape was analysed as described in the Methods. Box plots indicate the median (centre line) and the first and third quartiles (upper and lower hinges). **P < 0.01; Welch’s ANOVA followed by a Games–Howell post hoc test.

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