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. 2016 Apr 15;64(2):522–534. doi: 10.1002/hep.28550

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Copyright © 2016 The Authors. HEPATOLOGY 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 Creative Commons Attribution‐NonCommercial‐NoDerivs 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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Knockdown of AE2 induced sAC expression. AE2 was knocked down by lentivirus‐mediated shRNA interference in H69 cholangiocytes and confirmed by RT‐qPCR (A) and immunoblot (B). Expression of sAC messenger RNA (C) and protein level (D) in both short hairpin control and AE2 knockdown H69 cholangiocytes was examined. Immunoblots were quantified by ImageJ (E). Two‐tailed Student t test, *P < 0.05, **P ≤ 0.01. (F) Control and AE2KD H69 cholangiocytes were incubated with 50 μM etoposide or 0.1% dimethyl sulfoxide (vehicle) for 16 hours. Caspase 3/7 activity was determined. One‐way ANOVA, P < 0.0001. The figure shows a representative experiment from a series with similar results. ***P ≤ .001. Abbreviations: ATP1A1, adenosine triphosphatase, sodium/potassium transporting, alpha 1 polypeptide; KD, knockdown; n.s., not significant; SHC, short hairpin control.