Fig. 7: CCL17-dependent CCL3 release controls T_reg differentiation via CCR1.

a, Experimental scheme wherein CD4⁺CD62L⁺ T cells isolated from spleens of Apoe^−/−, Apoe^−/−Ccr1^−/− or Apoe^−/−Ccr5^−/− mice were cultured for 3 d under T_reg-polarizing conditions (TGFβ at 100 ng ml⁻¹) in the presence or absence of recombinant mouse CCL3 (100 ng ml⁻¹). b, Quantification of CD45⁺CD4⁺CD25⁺FoxP3⁺ T_reg cells (number of independent experiments per bar from left to right, n = 7, 7, 7, 7, 7, 7, 6, 6 and 6) using flow cytometry. c, Scheme of co-culture experiment wherein CD4⁺CD62L⁺ T cells isolated from spleens of Apoe^−/−, Apoe^−/−Ccr1^−/− or Apoe^−/−Ccr5^−/− mice were combined with sorted CD45⁺CD11c⁺MHCII⁺eGFP⁺ cDCs from LNs of Apoe^−/−Ccl17^wt/e or Apoe^−/−Ccl17^e/e mice and cultured for 3 d. d, Quantification of CD45⁺CD4⁺CD25⁺FoxP3⁺ T_reg cells (number of independent experiments per bar from left to right, n = 5, 5, 5, 4, 5 and 3) using flow cytometry. e, Experimental scheme of Apoe^−/− or Apoe^−/−Ccr1^−/− mice fed a WD for 12 weeks (e–j). f, Representative images and quantification of lesion area after ORO staining for lipid deposits in the aortic root of Apoe^−/− (n = 10) or Apoe^−/−Ccr1^−/− (n = 9) mice. Scale bar, 500 µm. g, Quantification of lesion area after ORO staining for lipid deposits in the thoraco-abdominal aorta of Apoe^−/− (n = 10) or Apoe^−/−Ccr1^−/− (n = 8) mice. h, Representative images and quantification of atherosclerotic lesion size in aortic arches of Apoe^−/− (n = 11) or Apoe^−/−Ccr1^−/− (n = 8) mice after H&E staining. i,j, Flow cytometric quantification of CD45⁺CD3⁺CD4⁺CD25⁺FoxP3⁺ T_reg cells in para-aortic LNs (i) and spleen (j) of Apoe^−/− (i,j, n = 13 each) or Apoe^−/−Ccr1^−/− (i, n = 8; j, n = 9) mice. Data represent mean ± s.e.m. (a–j). Two-sided P values as indicated and as analyzed by a mixed-effects regression model followed by pairwise contrasts for fixed factors corrected by a Holm–Šídák’s procedure (b), unpaired Student’s t-test (d,f,i) or Mann–Whitney U-test (g,h,j).

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