Fig. 2: CCL3 induction by CCL17 does not require CCR4 but inversely correlates with T_reg numbers.

a, Experimental scheme of Apoe^−/−, Apoe^−/−Ccl17^e/e and Apoe^−/−Ccr4^−/− mice fed a WD. b,c, CCL3 plasma concentrations in Apoe^−/− (n = 17) and Apoe^−/−Ccl17^e/e (n = 18) mice (b) or Apoe^−/− (n = 18) and Apoe^−/−Ccr4^−/− (n = 17) mice (c), as measured by ELISA. d, Experimental scheme of reconstituting irradiated Apoe^−/− or Apoe^−/−Ccl17^e/e mice with Apoe^−/− BM before WD feeding. e, CCL3 concentrations in plasma of Apoe^−/−►Apoe^−/− (n = 5) or Apoe^−/−►Apoe^−/−Ccl17^e/e (n = 7) mice were measured by ELISA. f,g, Sorted CD11c⁺MHCII⁺ cDCs (control, n = 17 replicates over 11 independent experiments; CCL17, n = 23 replicates over 12 independent experiments) (f) or CD3⁺ T cells (control and CCL17, n = 17 replicates over six independent experiments) were cultured for 4 h with or without recombinant mouse CCL17 (g). CCL3 concentrations in the supernatant were measured by multiplex bead array. h, Isolated T cell subsets from LN suspensions of Apoe^−/− mice were stimulated for 4 h in the presence or absence of CCL17. CCL3 concentrations in supernatants were measured by ELISA (n = 6 for each bar). i, Sorted CD11c⁺MHCII⁺eGFP⁻ cDCs (CCL17-competent, control and CCL17, n = 10) or CD11c⁺MHCII⁺ eGFP⁺ cDCs (CCL17-deficient, control n = 12; CCL17 n = 10) from Apoe^−/−Ccl17^e/WT mice were cultured for 4 h in the presence or absence of CCL17. CCL3 concentrations in supernatants were measured by ELISA. j, Sorted CD11c⁺MHCII⁺ cDCs from LNs of Apoe^−/− mice were cultured for 4 h in the presence or absence of CCL17 with or without the CCR4 inhibitor C021, and CCL3 concentrations in supernatants of isolated cDCs were measured by multiplex bead array (n = 20 replicates across six independent experiments for all conditions). k, Isolated T cell subsets from LN suspensions of Apoe^−/− mice were stimulated with or without CCL17 in the presence of absence of the CCR4 inhibitor C021 for 4 h and CCL3 concentrations in supernatants were measured by ELISA (control, n = 39 replicates; C021, n = 38 replicates; CCL17, 42 replicates; CCL17 + C021, n = 40 replicates; all distributed over nine independent experiments). Data represent mean ± s.e.m. (a–k). Two-sided P values as indicated versus control/Apoe^−/− and as analyzed by unpaired Student’s t-test (b,c,e), nested t-test (f,g), two-way analysis of variance (ANOVA) (h) or nested ANOVA (j,k) with Holm–Šídák’s post hoc test and Kruskal–Wallis H with Dunn’s post hoc test (i).

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