Figure 5
Tim-4 (T-cell immunoglobulin- and mucin-domain-containing molecule-4) inhibits cytokine production via nuclear factor-kappa B (NF-κB) signaling pathway. Tim-4 overexpression stable cell line RAW264.7-Tim-4 and control cells RAW264.7-pcDNA3 were pretreated with 83.3 μg/ml of NF-kB inhibitory ligand for 30 min. Lipopolysaccharide (LPS; 100 ng/ml) was used to stimulate these cells for 24 h, and tumor necrosis factor-α (TNF-α) (a) and interleukin-6 (IL-6) (b) levels in supernatants were measured by ELISA. Peritoneal macrophages were pretreated with 83.3 μg/ml of NF-kB inhibitory ligand for 30 min, then incubated with 5 μg/ml Tim-4 blocking antibody or isotype control IgG for 1 h. LPS (100 ng/ml) was used to stimulate these cells for 24 h, and TNF-α (c) and IL-6 (d) levels in supernatants were measured by ELISA.
