Fig. 7

Astrocyte-derived CCL2 and CCL9 but not CCL1 regulate macrophage polarization, in vitro. a Macrophage cultures were exposed to ODN 2088-treated astrocyte CM (ODN 2088-CM), in the absence or presence of CCL1 neutralizing Ab. The graph shows the quantification of the F4/80⁺/Arg-1⁺ cell number expressed as percentage of total F4/80⁺ cells in the macrophage cultures [p = 0.7228, independent-sample t-test, two-tailed]. b Macrophage cultures were exposed to vehicle-treated astrocyte CM, in the absence or presence of CCL2 neutralizing Ab. The graph shows the quantification of the F4/80⁺/Arg-1⁺ cell number expressed as percentage of total F4/80⁺ cells in the macrophage cultures [**p < 0.01, independent-sample t-test, two-tailed]. c Macrophage cultures were exposed to vehicle-treated astrocyte CM, in the absence or presence of CCL9 neutralizing Ab. The graph shows the quantification of the F4/80⁺/Arg-1⁺ cell number expressed as percentage of total F4/80⁺ cells in the macrophage cultures [***p < 0.001, independent-sample t-test, two-tailed]. d Macrophage cultures were exposed to vehicle-treated astrocyte CM (Veh-CM) or ODN 2088-treated astrocyte CM (ODN 2088-CM) for 24 h, with or without (control) addition of rmCCL9 (20 pg/ml). The graph shows the quantification of the F4/80⁺/Arg-1⁺ double-labeled cells expressed as percent of total F4/80⁺ cells in macrophage cultures [F (2, 6) = 53.68, p < 0.0001 by one-way ANOVA, *p < 0.05, **p < 0.01, ***p < 0.001 by Tukey’s post hoc test]. The experiments were independently repeated twice, yielding similar results. Results from a representative experiment are shown. Results obtained from additional biological repeats of these experiments can be found in Additional file 8D-G. Data are presented as mean ± SEM