Fig 5. Effects of tyrosol on intracellular calcium and the NF-κB activation in mast cells.

(A) RBL-2H3 cells (3 × 10⁴/well) were sensitized with anti-DNP IgE (50 ng/ml). After incubating overnight, the cells were preincubated with Fluo-3/AM for 1 h. The cells were pretreated with or without tyrosol for 1 h and then challenged with DNP-HSA (100 ng/ml). Intracellular calcium was detected every 1 min for 5 min using a fluorescent plate reader. Area under the curve (AUC) was calculated over 5 min. BAPTA, a calcium chelator, was used as a positive control. Each data presented as a graph represents the mean ± SE of three independent experiments. *Significant difference from DNP-HSA challenged group at P < 0.05. (B) RBL-2H3 cells (2 × 10⁶/well) were sensitized with anti-DNP IgE (50 ng/ml). After incubating overnight, the cells were pretreated with or without tyrosol for 1 h and then challenged with DNP-HSA (100 ng/ml). Activations of IKK complex and NF-κB were assayed by Western blot (N-: nuclear, p-: phosphorylated). The bands of actin and total form were used as a loading control. PP2, a Src family inhibitor, was used as a positive control. The band is a representative of three independent experiments.