Fig 5.

Tim-3 regulates IL-23p19 expression through phosphorylation of STAT3 in CD14⁺ cells cocultured with HCV⁺ hepatocytes. (A) Purified CD14⁺ monocytes were incubated with HCV^+/− Huh7 hepatocytes for 48 h, with or without TLR stimulation for 6 h, followed by flow cytometric analysis of Tim-3 and phosphorylated STAT3 expression in monocytes. Means + the SD from four repeated experiments are shown in the right panel. *, P < 0.05 (as determined by Student t test). (B) Western blot of STAT3 phosphorylation in purified monocytes incubated with HCV^+/− hepatocytes after TLR stimulation. Total STAT3 is probed to serve as a protein load control. Representative imaging and the means + the SD from four experiments with densitometry ratios of phosphorylated STAT3/total STAT3 protein in monocytes cocultured with HCV⁺ versus HCV⁻ hepatocytes are shown in the right panel. *, P < 0.05 (as determined by Student t test). (C) Purified CD14⁺ monocytes were incubated with HCV⁺ Huh7 cells with or without a STAT3-specific inhibitor for 48 h and stimulated with LPS/R848 for 6 h, followed by flow cytometric analysis. The results of representative experiments of IL-23p19 expression and phosphorylated STAT3 expression in monocytes, in the presence or absence of STAT3 inhibitor, are shown.