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. 2018 Sep 25;2(10):1247–1258. doi: 10.1002/hep4.1240

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© 2018 The Authors. Hepatology Communications published by Wiley Periodicals, Inc., on behalf of the American Association for the Study of Liver Diseases.

This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.

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CEACAM1 expression increased on Huh7.5.1 cells infected with JFH‐1 and decreased after treatment. (A) Expression of ligands for NK cells, such as BAT3, ICAM‐1, ICAM‐2, nectin‐2, PVR, MICA/B, ULBP1, ULBP2/5/6, ULBP3, HLA‐A, B, C, HLA‐E, HLA‐G and CEACAM1, was analyzed on Huh7.5.1 cells and Huh7.5.1 cells infected with JFH‐1 (Huh7.5.1/JFH‐1 cells). The dotted line represents Huh7.5.1 cells, the solid line represents Huh7.5.1/JFH‐1 cells, and the shaded area represents the isotype control. (B) CEACAM1 expression on Huh7.5.1 cells and Huh7.5.1/JFH‐1 cells was analyzed by flow cytometry (left panel, means ± SD, n = 4). CEACAM1 levels in the supernatants of Huh7.5.1 cells and Huh7.5.1/JFH‐1 cells were analyzed by ELISA (middle panel, means ± SD, n = 4). In addition, levels of the CEACAM1 mRNA in Huh7.5.1 cells and Huh7.5.1/JFH‐1 cells were analyzed by qRT‐PCR (right panel, means ± SD, n = 4). (C,D) CEACAM1 levels at the cell surface and in supernatants of Huh7.5.1/JFH‐1 cells were measured by flow cytometry and ELISA, respectively, at the indicated times after infection (means ± SD, n = 4). (E) Immunofluorescence staining for NS5A (green) and 4’,6‐diamidino‐2‐phenylindole (DAPI; blue) was performed in Huh7.5.1/JFH‐1 (left panel) cells and Huh7.5.1/JFH‐1 cells treated with BILN (right panel). Levels of JFH‐1 mRNA in Huh7.5.1/JFH‐1 cells before and after treatment were evaluated by qRT‐PCR (means ± SD, n = 4). (F) Analysis of Huh7.5.1 cells, Huh7.5.1 cells + BILN, Huh7.5.1/JFH‐1 cells, and Huh7.5.1/JFH‐1 cells + BILN. CEACAM1 expression on these cells was analyzed by flow cytometry (means ± SD, n = 4). Levels of CEACAM1 mRNA in these cells were evaluated by qRT‐PCR (means ± SD, n = 4). ^* P < 0.05.