Fig. 7: Thymic stromal lymphopoietin (TSLP) induces CUL5-mediated inhibition of antiviral immunity.

a Dot graph showing the top 20 enriched pathways in the alveolar macrophages (AMs) of HDM-treated mice compared with those in PBS-treated mice, with viral infection-related pathways highlighted in red. Dot size represents the number of differentially expressed signals. Rich factor represents the ratio of the number of differentially expressed genes (DEGs) located in the pathway entries to the total number of all annotated genes located in the pathway target genes. p-value was calculated by Fisher’s exact test. b Immunoblotting of O-GlcNAc transferase (OGT) and CUL5 expression in lysates of bone marrow-derived macrophages (BMDMs) stimulated by 20 μM TSLP, 20 μM IL-33, 20 μM IL-4, and 100 μM IL-33 for indicated times. c Co-immunoprecipitation (Co-IP) and immunoblotting of BMDMs treated by 20 μM TSLP for 4 h. d Confocal microscopic imaging of BMDMs treated by 20 μM TSLP for indicated times. CUL5, red; OGT, green. Nuclei were stained with DAPI (blue). Yellow in merge indicates the co-localization of CUL5 and OGT. Scale bars, 5 μm. e Pearson’s correlation coefficient (PCC) was determined to evaluate the correlation of voxel intensity between the CUL5 (red) and OGT (green) channels in (d). f The fluorescence intensity on the white lines in (d) was calculated. g Immunoblot analysis of OGT ubiquitination in BMDMs treated with 20 μM TSLP for 4 h. h Immunoblot analysis of the indicated proteins in lysates of BMDMs stimulated by 4 μg/ml Poly(I:C) and pretreated with 20 μM TSLP with/without 10 μM MLN4924. i Heatmap summarizing Ifnb, Cxcl10, and Isg15 mRNA expression in BMDMs stimulated by 4 μg/ml Poly(I:C) and pretreated with 20 μM TSLP with/without 10 μM MLN4924. j ELISA was performed to determine IFN-β expression in BMDMs stimulated by 4 μg/ml Poly(I:C) and pretreated with 20 μM TSLP with/without 10 μM MLN4924. Data in (b)–(d) and (g, h) are representative of three independent experiments. Data represent the means ± s.e.m., n = 3 biological replicates. p-values were calculated by one-way ANOVA (Tukey’s test) in (e), or two-way ANOVA (Tukey’s test) in (j). Source data are provided as a Source Data file.