Figure 1

Construction of IgE-mediated mast cell degranulation TCRPs.

(a) The real time cellular analyzer xCELLigence captured a characteristic peak induced by DNP-BSA. RBL-2H3 cells were seeded overnight and then sensitized with IgE for 24 hours, before the stimulation of DNP-BSA. (b)20 k cells per well was the optimal condition for cell culture. Initial inoculums of 5 k, 10 k, 20 k, 40 k, 80 k cells per well were introduced in a 96-well E-plate. Fixed concentration of stimuli was added. (c) The reaction to DNP-BSA followed a dose-dependent manner. 20 k cells per well were seeded and treated with different concentrations of DNP-BSA. (d–f) Mast cell degranulation was coupled with mediators release and morphological dynamics. RBL-2H3 cells were sensitized with 100 ng/mL IgE and activated by 100 ng/mL DNP-BSA. The percentage of released β-hexosaminidase was measured at the indicated time. mRNA levels of cytokines and chemokines were evaluated by qPCR. Morphological changes were detected with LSFM as described in methods. Red: F-actin, blue: nucleus, scale bar = 25 μm. Data are mean ± SD and are representative of three independent experiments. *P < 0.05; **P < 0.01; ***P < 0.001.