Effect of GCSE treatment on T cells and B cells isolated from AD-induced mice. (A) Mouse splenocytes were incubated with various concentrations of GCSE for 72 hrs. Cell viability was estimated with WST-1 assay. (B) Draining lymph node CD19+ B cells isolated from AD-induced mice were stimulated with LPS (10 μg/ml)/IL-4 (5 ng/ml) in the presence of various concentrations of GCSE dissolved in 70% alcohol for 72 hrs. Mouse IgE levels in the supernatant of B cell culture were measured by ELISA. Same volume of 70% alcohol was treated as control. (C) Draining lymph node CD4+ T cells from AD-induced mice were stimulated with PMA (50 ng/ml)/ionomycin (1 μM) in the presence of GCSE (0.25 mg/ml) for 4 hrs. Relative expression of cytokines of GCSE treated samples was compared with control samples by qRT-PCR. Expression level of HPRT was used as an internal control. (D) CD4+ T cells from AD-induced mice were stimulated with PMA /ionomycin for 72 hrs in the presence of GCSE (0.25 mg/ml), then protein level of each cytokine was analyzed by ELISA. Error bars indicate SD. One (*), two (**) and three (***) indicate p < 0.05, p < 0.01, and p < 0.001 respectively. Data are representative of three independent experiments.