Fig. 3

The transcription factor c-Fos/AP-1 inhibitor T-5224 shows its anti-allergic effects on IgE/Ag activated RBL-2H3 cells. A Chemical structure of T-5224. B RBL-2H3 was incubated with T-5224 (0.1–50 μM). After 24 h, cell viability was measured by performing the CCK-8 assays. C Dual luciferase reporter assays performed to detect the selective inhibitory effects on AP1 signaling pathway in MCs. D IgE-sensitized RBL-2H3 were pre-incubated for 1 h with indicated concentration of T-5224 and then stimulated with DNP-HSA for 30 min; β-Hexosaminidase released into the supernatant was measured. E, F The gene expression of pro-inflammatory cytokines IL-4 and Egr 1 was analyzed 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 **p < 0.01 vs. nontreated activated cell