Fig. 6.

CCL20 antibody partly inhibits FOXO1 stimulation of PBMC migration in the presence of TNF-α. A, Effects of FOXO1 on PBMC migration. HepG2 were incubated in six-well plates and transfected with pcDNA3.1 (0.4 μg), FOXO1-AAA plasmid (0.4 μg), siFOXO1 (20 nmol/ml), or siNC (20 nmol/ml). Cells were treated 24 h after transfection with 10 ng/ml TNF-α or vehicle PBS for an additional 24 h, and the cell culture supernatant were used for migration test of monocyte-deprived PBMC (mainly lymphocytes) in the six-well Boyden chamber system. B, Effects of CCL20 antibody on PBMC migration. HepG2 cells were transfected with pcDNA3.1 or FOXO1-AAA plasmid and treated with TNF-α as described above in the presence of anti-human CCL20 antibody (10 ng/ml) or IgG. The supernatants were harvested for PBMC migration test. DMEM and DMEM plus purified human CCL20 protein (1 ng/ml) were used as negative and positive controls, respectively. Results were from five high-power fields (HPF) of each of triplicate wells. *, P < 0.05; and **, P < 0.001 vs. control.