8,9-EET sensitizes AITC induced TRPA1 response. (A) Representative calcium imaging experiment of 8,9-EET dependent TRPA1-sensitization. DRG cells were stimulated with AITC twice (50 μM, 15 sec) with 10 minutes interval. 1 μM 8,9-EET or vehicle was perfused for two minutes prior to the second AITC stimulation. (B) Statistical analysis comparing the amplitudes of AITC induced TRPA1 response. Data shown represent the mean ± SEM of 21-23 cells. Student's t test: *, p ≤ 0.05 (C) 8,9-EET sensitizes AITC induced CGRP release from sciatic nerve axons. Freshly isolated sciatic nerves were incubated with 10 μM 8,9-EET (ipsilateral) or 3.2% EtOH (vehicle, contralateral) and stimulated with 50 μM AITC. CGRP was determined by ELISA from extracellular solutions. Data shown represent the mean ± SEM from nerves of 3-4 animals. Two way ANOVA with Bonferroni post test: *, p ≤ 0.05.