Recombinant galectin-9 synergizes microglial TNF and IL-6 production. (A) Primary mouse microglia were unstimulated (CTL) or stimulated with recombinant mouse galectin-9 (2 μg/ml), poly(I:C) (25 μg/ml), or galectin-9 and poly(I:C) together for 24 hours and cell morphology examined by microscopy (A). (B-F) Following the above treatments chemokines and cytokines were simultaneously measured from supernatants by Bioplex assay. Levels of TNF (B), IL-6 (C), IL-1β (D), IL-12(p40) (E), CCL2 (F) and IL-10 (G) are shown. Results are means ± SE of triplicate samples and are representative of three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001; † P < 0.05 versus control. (H-J) Primary mouse microglia were stimulated as above for 24 hours, gated on CD11b (H) and the surface expression of CD86 (I) and MHC II (J) determined by flow cytometry. Representative histograms (left) as well as combined mean fluorescence intensity values from three independent experiments (right) are shown. Results are combined means ± SE of three independent experiments.