Fig. 7.

Upregulation of ICAM-1 in cholangiocytes in response to IFN-γ affects adherence of cocultured T cells. A and B: functional manipulation of miR-221 influenced adherence of H69 by cocultured Jurkat cells in an ICAM-1-dependent manner. H69 cells were first treated with miR-221 precursor (A) or anti-miR-221 (B) for 72 h. After being washed, cells were incubated with the medium containing the IgG isotype control (Ctrl) or anti-ICAM-1 mAb for 1 h. Activated Jurkat cells were labeled with the calcein acetoxymethyl ester (calcein AM) and then incubated with H69 cells. Adherence of Jurkat cells was measured under the fluorescence microscope and presented as percentage of control. C: IFN-γ stimulation increased adherence of H69 cells by cocultured Jurkat cells. H69 cells were exposed to IFN-γ for 24 h followed by treatment with the anti-ICAM-1 mAb or IgG isotype for 1 h. H69 cells were treated with miR-221 precursor for 72 h before exposure to IFN-γ, and adherence of H69 cells by Jurkat cells was then determined. Data are representative of 3 independent experiments. Precursor-Ctrl, nonspecific precursor control. *P < 0.05 t-test vs. IgG isotype control (in A and B) or non-IFN-γ-stimulated cells (in C); #P < 0.05 t-test vs. precursor-Ctrl or IgG isotype control. D: effects of upregulation of ICAM-1 in H69 cells in response to IFN-γ on adherence of cocultured T cells as assessed by immunofluorescent microscopy. Calcein AM-labeled Jurkat cells following adherence to H69 cells were shown. Bars = 50 μm.