Fig. 2.

Localization of TRPA1 and TRPV3 in HBECs. (A) Functional expression of TRPA1 and TRPV3 in lobar HBECs using a calcium flux assay. Agonists of TRPA1 (AITC, 300 μM), TRPA1/TRPV3 (pine WSPM, 78 μg/cm², and carvacrol, 250 μM), and TRPV3 (drofenine, 250 μM) were used in combination with either the EGTA (50 μM) and ruthenium red (250 μM) to block the influx of extracellular calcium into cells, or thapsigargin (Thaps.; 2.5 μM) to deplete intracellular/ER calcium stores. Calcium flux data (over a 100-second period) are represented as a percentage of the maximum fluorescence in the cells, elicited by the calcium ionophore ionomycin (10 μM). Data are represented as means ± S.D. from n = 3 replicates. *P < 0.05; ***P < 0.001; ****P < 0.0001 using ordinary two-way ANOVA and the Tukey post-test comparing each treatment to the respective agonist only control. (B) Representative immunostaining of TRPV3 (green), the ER biomarker calnexin (red), and nuclei (blue) in lobar HBECs. The complete fluorescence micrographs may be found in Supplemental Figure 3.