Fig. 1: CCR4 does not affect atherosclerosis or T_reg cells, while CCL17-deficiency increases tolerogenic DCs.

a, Experimental scheme of WD feeding. b, Representative sections and quantification of lesion area after Oil-Red-O staining (ORO) for lipid deposits in the aortic root of Apoe^−/− (n = 16) or Apoe^−/−Ccr4^−/− (n = 20) mice. Scale bar, 500 µm. c, Quantification of lesion area after ORO in the thoraco-abdominal aorta of Apoe^−/− (n = 5) or Apoe^−/−Ccr4^−/− (n = 19) mice. d, Atherosclerotic lesion size quantified by hematoxylin and eosin (H&E) staining in the aortic arches of Apoe^−/− (n = 16) or Apoe^−/−Ccr4^−/− (n = 19) mice. e,f, Representative dot plots and flow cytometric quantification of CD45⁺CD3⁺CD4⁺CD25⁺FoxP3⁺ T_reg cells in para-aortic LNs (e) and spleen (f) of Apoe^−/− (e, n = 16; f, n = 16) or Apoe^−/−Ccr4^−/− (e, n = 19; f, n = 20) mice. g, Co-culture of CD45⁺CD11c⁺MHCII⁺ DCs sorted from Apoe^−/−, Apoe^−/−Ccl17^e/e or Apoe^−/−Ccr4^−/− mice with splenic CD4⁺CD62L⁺ T cells isolated from Apoe^−/− or Apoe^−/−Ccr4^−/− mice. After 72 h, the abundance of CD45⁺CD3⁺CD4⁺CD25⁺FoxP3⁺ T_reg cells was determined by flow cytometry (individual number of experiments per bar from left to right n = 7, 5, 4, 6 and 5). Besides the cells, no other factors were added to the culture. h–j, CD45⁺CD3⁻CD19⁻CD11c⁺ cells were sorted from pooled LNs of Apoe^−/−Ccl17^wt/e or Apoe^−/−Ccl17^e/e mice on chow diet. Uniform Manifold Approximation and Projection (UMAP) projection of 4,731 single cells colored by inferred cell types consisting of seven distinct DC clusters (h). Depicted are eGFP⁺ cell counts in CCR7⁺ DCs or other (CCR7⁻) DCs (percentage of total numbers) and proportions of four distinct CCR7⁺ DC clusters among all single cells (CD45⁺CD3⁻CD19⁻CD11c⁺) (bottom) (i). j,k, Tolerogenic score calculated for four distinct CCR7⁺ DC clusters and other (CCR7⁻) DC clusters based on the top 20 genes differentially expressed between the groups (top) as (1 + mean (top 20 upregulated tolerogenic genes))/(1 + mean(top 20 upregulated immunogenic genes)) (j). k, Table depicts cell counts of four CCR7⁺ DC and other DC clusters. LNs for scRNA-seq were obtained from Apoe^−/−Ccl17^wt/e (n = 8) and Apoe^−/−Ccl17^e/e (n = 6). Data represent mean ± s.e.m. (a–k). Two-sided P values as indicated and as analyzed by unpaired Student’s t-test (b,e,f), Mann–Whitney U-test (c,d) or Kruskal–Wallis H with Dunn’s post hoc test (g,j). Differences in proportion in DC clusters between Apoe^−/−Ccl17^wt/e and Apoe^−/−Ccl17^e/e were computed by Pearson’s chi-squared test (k).

Source data