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. 2022 May 25;41:182. doi: 10.1186/s13046-022-02401-6

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© The Author(s) 2022

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 2 — TROY enhances the stemness properties of HCC cells. (A, B) Western blotting of TROY (A), NANOG, SOX2, and OCT3/4 (B) in 8024 and Hep3B HCC cells transfected with lentiviruses expressing empty vector (vec) or TROY isoforms (#1 and #2). β-Actin was used as a loading control. (C) Spheroid formation assay (left) and summaries (right) of spheroid numbers in 8024 and Hep3B cells with vec or TROY isoforms. (D) Representative flow cytometry plots (left) and summary (right) of apoptotic cells in 8024 and Hep3B cells transfected with vec or TROY isoforms. Cells were treated with Sorafenib (20 μm) for 24 h. (E) Representative flow cytometry plots (left) and summary (right) of ALDH activity in 8024 and Hep3B cells transfected with vec or TROY isoforms. (F) Tumor (left), tumor volume (middle) and tumor weight (right) of in vivo tumor formation assays with subcutaneously injected Hep3B cells transfected with vec or TROY isoforms in NOD/SCID mice. (G) In vivo subcutaneous implantation nude mice model induced by 8024 cells transfected with vec or TROY isoforms. Statistical significances: *, P < 0.05; **, P < 0.01; ***, P < 0.001