Figure 2.

TRPV₁ expression in children’s HBE cells infected by RSV. HBE cells from children with and without asthma were infected with RSV (MOI = 1) and assessed for TRPV₁ expression at 0–24 h post infection from children without (2, 9) and with asthma (14, 15) (A) or 0, 3, and 6 h post infection from children without (2, 6, 7, 10) and with asthma (12–15) (B). TRPV₁ protein expression was measured by Western blot and normalized to actin. C: quantification of results from (A); n = 3 independent experiments. D: representative fluorescent micrographs of immunoreactive TRPV₁ (green) expression in permeabilized HBE cells from children without (2, 8, 10) and with asthma (12, 13) at 0, 3, and 6 h after infection with RSV (MOI = 1). E: quantification of results from C, showing densitometry measurements on a minimum of 15 cells/donor. F: TRPV₁ (green) expression in plasma membranes (PMs, red) of nonpermeabilized HBE of children without (2, 8, 10) and with asthma (12–14) at 0, 3, and 6 h post infection with RSV. White arrowheads indicate the plasma membranes. G: quantification of results from F showing TRPV_1PM calculated as the average of densitometric measurements on a minimum of 15 cells/donor. Scale bar = 20 µm. H: TRPV₁ expression in the biotinylated plasma membrane fraction of HBE cells from children without (2, 4, 7) and with asthma (12, 13, 15) incubated with sterile medium or RSV (MOI = 1) for 6 h. Cells were labeled with sulfo-NHS-biotin and purified with streptavidin. Input lysates are shown in the lower bands. I: quantification of the results from G normalized to flotillin. Data are expressed as means ± SE and were analyzed using the Kruskal–Wallis test with Dunn’s multiple comparisons. *P < 0.05, ***P < 0.001, ****P < 0.0001 compared with the 0-h time point; #P < 0.05, ##P < 0.01, ###P < 0.001 compared with nonasthmatic controls. HBE, human bronchial epithelium; MOI, multiplicity of infection; RSV, respiratory syncytial virus; TRPV1, transient receptor potential vanilloid 1.