Fig. 3

IgE-IC elicits acute ocular itch in naive mice. A Timeline of behavioral recording, drug application, and tissue collection. B Mice were ocular instilled with IgE-IC (1, 10, 50 μg/ml; 5 μl), monomeric IgE (50 μg/ml; 5 μl), OVA (100 mg/ml, 5 μl) or vehicle (PBS; 5 μl), and eye-towards scratching bouts were counted over 1–12 h. Baseline was identified as recorded without any instillation. n = 8–10 mice per group; **P < 0.01 vs. PBS, ***P < 0.001 vs. PBS, ****P < 0.0001 vs. PBS; ^###P < 0.001 vs. Baseline, ^####P < 0.0001 vs. Baseline; 2-way ANOVA for repeated measures followed by Bonferroni’s post hoc test. C Representative images of conjunctiva taken 1 h after ocular instillation with PBS, monomeric IgE (50 μg/ml), or IgE-IC (50 μg/ml), stained with H&E. Scale bar: 100 μm. D–F Representative images of conjunctiva taken 1 h after ocular instillation with either PBS, monomeric IgE (50 μg/ml), or IgE-IC (50 μg/ml) and stained for Ly6C/G (D), IBA1 (E), and CD3 (F). Scale bar: 100 μm. The embedded boxes showed the enlarged views. G Quantification for fluorescence intensity of markers showed no significant differences between treatment groups. n = 4 per group; one-way ANOVA followed by Bonferroni’s post hoc test comparisons. H Quantitative analysis of Ly6C/G+, IBA+, and CD3+ cell numbers after different treatments. n = 4 per group; one-way ANOVA followed by Bonferroni’s post hoc test comparisons. I Representative images of conjunctiva taken 1 h after ocular instillation with either PBS, monomeric IgE (50 μg/ml), or IgE-IC (50 μg/ml) and stained for FITC-avidin. Scale bar: 100 μm. The embedded boxes showed the enlarged views. J Quantitative analysis of mast cells number after different treatments. n = 4 per group; one-way ANOVA followed by Bonferroni’s post hoc test comparisons. K Proportions of activated mast cells (with degranulation) after different treatments. n = 4 per group; one-way ANOVA followed by Bonferroni’s post hoc test comparisons