Supplementary material

Supplementary material

Please note: supplementary material is not edited by the Editorial Office, and is uploaded as it has been supplied by the author.

  • Supplementary materials and methods
  • Supplementary figure S1 - TDI activates murine TRPA1-expressing CHO cells: Calcium imaging. Intracellular Ca2+ imaging experiments show the effect of extracellular applications of 0.001% TDI or TDA on murine TRPA1 (a) and TRPV1 (b), transfected in CHO cells, respectively. Solid lines represent the means and dashed lines represent the corresponding SEM. Cells responding to 0.001% TDI and AITC or CAP, and 0.001% TDA and AITC or CAP are represented in a black and red line, respectively, while unresponsive cells are represented in blue lines. Bars represent percentages of responding cells to TDI (black) and TDA (red).
  • Supplementary figure S2 - Figure 2. Individual responses in the methacholine tests of all experimental groups. Each symbol represents the methacholine reactivity of an individual mouse, expressed as the AUC of Rn to methacholine (from 0 to 20 mg/ml) (as shown in fig. 2a, 3a, 4a, 5a, 5b, 6a, 6b, 6c, 7a, 8a and online supplementary figure S5a); horizontal lines are group means, n=5–9 per group. **p<0.01 and ***p<0.001 compared to own control group.
  • Supplementary figure S3 - Figure 3. Involvement of TRPA1 in the development of TDI-induced AHR: pharmacological blockade. Total serum IgE levels in WT mice treated with either TRPA1 antagonist or vehicle (a) or either NK1R antagonist or vehicle (c). Number of Th- (CD4+), Tc- (CD8+) and B-lymphocytes (CD19+) for TRPA1 (b) or NK1R (d) antagonist. Cytokine release of IL-13, IFN-γ and IL-10 for NK1R antagonist (e). Data are represented as mean±SD, n=6–12 per group. *p<0.05, **p<0.01 and ***p<0.001 compared to own control group.
  • Supplementary figure S4 - Figure 4. Induction of AHR depends on an allergen-specific challenge. Total serum IgE levels (a). Number of Th- (CD4+), Tc- (CD8+) and B-lymphocytes (CD19+) (b-d). Data are represented as mean±SD, n=6–10 per group. **p<0.01 and ***p<0.001 compared to own control group (AOO/AITC, AOO/CAP or AOO/AITC/AOO).
  • Supplementary figure S5 - Figure 5. Induction of lymphocytes in the development of TDI-induced AHR. Mean values of Rn in Rag2 KO mice (a). Total serum IgE levels are represented as mean±SD (b). n=5–6 per group. ***p<0.001 compared to Rag2 KO AOO/AOO control group.
  • Supplementary figure S6 - Figure 6. Involvement of mast cells in the development of TDI-induced AHR. Total serum IgE levels (a). Number of Th- (CD4+), Tc- (CD8+) and B-lymphocytes (CD19+) (b) and ex vivo cytokine release of IL-13, IFN-γ and IL-10 (c). Data are represented as mean±SD, n=5–6 per group. *p<0.05, **p<0.01 and ***p<0.001 compared to AOO/ Veh/AOO control group.