Fig. 1. Notch4 expression on lung T_reg cells in allergic airway inflammation.

a, RT-PCR of Notch1-4 transcripts in lung T_reg and T_eff cells isolated from PBS, OVA and OVA+UFP mouse groups (n=5). b,c, Flow cytometric analysis, cell frequencies and mean fluorescence intensity (MFI) of Notch4 expression on lung T_reg and T_eff cells in the respective treated groups (n=15). d, Flow cytometric analysis and cell frequencies of Notch4 expression on OT-II⁺CD4⁺Foxp3⁺ T cells generated in co-cultures with sham or OVA_323–339+UFP-pulsed alveolar macrophages without or with IL-6 or anti-IL-6R mAb (n=5). e, Flow cytometric analysis and cell frequencies of Notch4⁺Helios^– and Helios⁺ lung T_reg cells isolated from the respective treated groups (n=5). f, Flow cytometric analysis and cell frequencies of Notch4 expression on in vitro differentiated T_reg cells derived from naive CD4⁺ T cells isolated from Foxp3^YFPCre, Foxp3^YFPCreIl6r^Δ/Δ and Foxp3^YFPCreStat3^Δ/Δ mice and either untreated or treated with IL-6 (n=6). g, ChIP assays for the binding of STAT3 and control (IgG) antibodies to the Notch4 promoter in lung T_reg cells of OVA+UFP-treated Foxp3^YFPCre, and Foxp3^YFPCreStat3^Δ/Δ mice (n=6). Each symbol represents one mouse. Numbers in flow plots indicate percentages. Error bars indicate SEM. Statistical tests: One-way ANOVA with Dunnett’s post hoc analysis (b,c,f); two-way ANOVA with Sidak’s post hoc analysis (a,d,e,g). *P<0.05, ***P<0.001, ****P<0.0001. Data representative of two or three independent experiments.