Fig 2: Expression of TSLP on cardiovascular tissue. A and D, Representative images of TSLP staining in the heart and the aorta. B and E, Quantitative PCR analysis of TSLP gene expression in the heart and the aorta (median with 25th and 75th percentiles; Kruskal–Wallis test; #P=0.0915, **P=0.0013). C and F, Representative images and intensity analysis of Western blot for the levels of cardiac (***P=0.0007) and aorta TSLP protein (median with 25th and 75th percentiles; Kruskal–Wallis test; #P=0.9999; **P=0.0022) from each treatment. Data are shown as the average±SD of 3 to 6 independent experiments. Black bar=100 μm. ****P<0.0001. ND indicates normal diet; PCR, polymerase chain reaction; SD, standard deviation; TSLP, thymic stromal lymphopoietin; WTD, Western‐type diet.

Fig 3: Coculture of BM‐DCs and T cells. A, Representative FACS analysis of the percentages of nTregs and CD4+LAP+ Tregs in CD4+ splenic T cells after 7 days of culturing. B and C, Graphs represent the average percentages of nTregs and CD4+LAP+ Tregs in CD4+ splenic T cells. D, Quantification of IFN‐γ (#P=0.0578 vs no DCs; #P=0.8652 vs iDCs) and IL‐4 released by CD4+ T cells. E and F, Proliferative response of purified CD4+ (#P=0.5676, *P=0.0163) or CD4+CD25+ T cells exposed to different stimulus. G, Conversion of naive CD4+CD25− T cells into CD4+CD25+Foxp3+ T cells under different conditions (**P=0.0019; ***P=0.0003). Data are shown as the average±SD of 3 to 5 independent experiments. ****P<0.0001. APC indicates allophycocyanin; BM, bone marrow; DCs, dendritic cells; iDCs, immature DCs; FITC, fluorecein isothiocyanate; IFN, interferon; IL, interleukin; LAP, latency‐associated peptide; oxLDL, oxidized low‐density lipoprotein; PE, phycoerythrin; SD, standard deviation; TSLP, thymic stromal lymphopoietin.

Fig 4: TSLP-DCs attenuate the development of atherosclerosis (?). A, Representative sections of aortic sinus stained with Oil Red O and antibodies specific for MOMA-2 for macrophages, Foxp3 for Tregs, a-smooth muscle actin staining, and collagen staining in ApoE-/- mice in different groups (no DCs, iDCs, oxLDL-DCs, and TSLP-DCs) at 16 weeks. Black bar=200 or 100 µm (Foxp3); black arrows indicate examples Foxp3-positive cells. B through F, Quantitative analysis of lesion size (**P=0.0074 vs oxLDL-DCs; **P=0.0047 vs TSLP-DCs), MOMA-2 (#P=0.6304; *P=0.0391), Foxp3 (median with 25th and 75th percentiles; Kruskal–Wallis test; *P=0.0158), a-SMA (#P=0.9994 vs TSLP-DCs; #P=0.9647 vs oxLDL-DCs), and collagen (#P=0.9878 vs TSLP-DCs; #P=0.9878 vs oxLDL-DCs) staining in different groups. Values are presented as the mean±SD, n=7 to 9. DCs indicates dendritic cells; iDCs, immature DCs; MOMA, monocyte/macrophage; oxLDL, oxidized low-density lipoprotein; SD, standard deviation; SMA, smooth muscle actin; TSLP, thymic stromal lymphopoietin.

Fig 5: Effects of TSLP-DC administration on nTregs, CD4+LAP+ Tregs, and Th1, Th2, and Th17 cells in spleens in ApoE-/- mice on day 14. A, CD4+ T-cell subsets were gated (a). Representative pictures of nTregs (b), CD4+LAP+ Tregs (c), Th1 (d), Th2 (e), and Th17 (f) are shown. Numbers in upper right quadrants indicate percentages of these cells. B, Results of statistical analysis of average percentages of nTregs (median with 25th and 75th percentiles; Kruskal–Wallis test; *P=0.0374 vs no DCs; #P=0.2596 vs iDCs; *P=0.0162 vs oxLDL-DCs) and Th1 cells (**P=0.0058; ***P=0.0007) in different groups. C, Results of statistical analysis of average percentages of CD4+LAP+ Tregs (**P=0.0010 vs no DCs; **P=0.0012 vs iDCs; ***P=0.0008), Th2 (median with 25th and 75th percentiles; Kruskal–Wallis test; #P=0.9999; *P=0.0226; **P=0.0066), and Th17 cells (#P=0.9726) in different groups. Six mice were analyzed for each group. Values are presented as the mean±SD. ****P<0.0001. APC indicates allophycocyanin; DCs, dendritic cells; iDCs, immature DCs; IFN, interferon; IL, interleukin; LAP, latency-associated peptide; oxLDL, oxidized low-density lipoprotein; SD, standard deviation; TSLP, thymic stromal lymphopoietin.