Fig. 4

T-5224 attenuates IgE-mediated BMMCs activation. A RBL-2H3 was incubated with T-5224(0.1–50 μM). After 24 h, cell viability was measured by performing the CCK-8 assays. IgE-sensitized BMMCs were pre-incubated for 1 h with indicated concentration of T-5224 and then stimulated with DNP-HSA for 30 min; β-hexosaminidase (B) and histamine (C) released into the supernatant was measured. D The gene expression of inflammatory cytokine IL-4 was determined by RT-PCR. E The secretion of inflammatory cytokines IL-4 was measured by ELISA. F The gene expression of Egr 1 was determined by RT-PCR. G, H Total protein was extracted, and the protein samples were analyzed by western blotting using specific antibodies (Egr 1, ERK, p-ERK, JNK, p-JNK, p38, p-p38). The band shown is representative of three independent experiments. Band signals (left panel) were analyzed by Image J software and the fold increase of signal levels compared with the Ctrl group was measured (right panel). Ctrl: control group without DNP-HSA treatment. Data summarize three independent experiments; *p < 0.05 vs. nontreated activated cell