Fig. 8
IgE upregulates neuronal FcεRIα and enhanced neuronal response to IgE-IC. A qRT-PCR revealed that FcεRIα mRNA was not changed by IgE. One-way ANOVA followed by Bonferroni’s post hoc test comparisons, n = 4. B Representative Western blotting gel bands of FcεRIα protein from dissociated TG neurons cultured with IgE of different concentrations. Western blotting revealed a significant increase in the protein levels of FcεRIα in dissociated TG neurons cultured with IgE. *P < 0.05, **P < 0.01, vs. Control, one-way ANOVA followed by Bonferroni’s post hoc test comparisons, n = 4. C IgE upregulated the FcεRIα protein in dissociated TG neurons as blocking protein synthesis by Cycloheximide (CHX). *P < 0.05, CHX vs. CHX+IgE, Student’s t-test, n = 3. D TG neurons cultured with (right, IgE group) and without (left, Control group) IgE (5 μg/ml) were exposed to IgE-IC (IC; 0.1 μg/ml; 30 s), followed by capsaicin (CAP; 1 μM; 10 s) and KCl (50 mM; 10 s). Neuronal activation was analyzed by calcium imaging (representative trace). E Proportion of responsive small-diameter neurons as IgE-IC application was significantly increased in IgE group compared with Control group. *P < 0.05, IgE vs. Control, χ2 test. F Mean magnitude of IgE-IC-induced R (340/380) growth was also elevated in IgE group. ***P < 0.001, IgE vs. Control, Student’s t-test. G Protein levels of FcεRIα in TG from mice sensitized different times were determined by Western blotting. #P < 0.05, ##P < 0.01 vs. Control; *P < 0.05 vs. Sensitized 1w, one-way ANOVA followed by Bonferroni’s post hoc test comparisons. H Ocular scratching behavior as challenged by OVA (1% in PBS) or vehicle in mice sensitized different times. n = 6 per group, ***P < 0.001, ****P < 0.0001 vs. Control; ##P < 0.01, ####P < 0.0001 vs. Sensitized 1w; ^^^P < 0.001 vs. Sensitized 2w; Student’s t-test