Figure 5

Tim-3 is not required for the effects of galectin-9 on microglial cytokine production. (A) RNA was extracted from rat microglia or astrocytes. Expression of Havcr3 (top), the microglia marker Iba-1 (middle), and the astrocyte marker Gfap (bottom) were examined by RT-PCR. (B-C) Surface expression of Tim-3 was examined on CD11b⁺ primary mouse microglia (B; left) and BV2 cells (C; right) by flow cytometry. Results are representative of three independent experiments for each cell type. (D-E) The effect of Tim-3 neutralization on TNF production in mouse mixed glial cultures. (D) Cultures were stimulated with poly(I:C) for 24 hours in the presence of increasing concentrations of a neutralizing antibody to Tim-3 (8B.2C12) or an isotype control (rat IgG1). (E) Cultures were stimulated with poly(I:C) and increasing concentrations of soluble Tim-3. Results are means ± SE of duplicate samples and are representative of three independent experiments for each experiment. (F) Mouse BV2 cells were challenged with vehicle, LPS (100 ng/ml) or poly(I:C) (25 μg/ml) in the presence of increasing concentrations of recombinant mouse galectin-9 (2 μg/ml) and TNF levels measured after 24 hours. Results are means ± SE of duplicate samples and are representative of two independent experiments.