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. 2021 May 25;11:10931. doi: 10.1038/s41598-021-90263-x

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© The Author(s) 2021

Open Access This 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/.

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Reprogramming of liver fibrosis in the multicellular tumor spheroid (MCTS) model but not the multicellular hepatocyte spheroid (MCHS) model. (A) Structure through F-actin staining of MCTS treated with 5 µM of retinoic acid or forskolin. (B) Expression of mesenchymal-related marker (α-SMA), and an EndMT-related marker (CD31) in tumor spheroids or MCTSs with or without treatment with 1 µM retinoic acid or forskolin. (C) Expression of cancer stem cell-related marker (CD133) in MCTS with or without 1 µM retinoic acid or forskolin (left panel). The expression of CD133 was quantified (right panel). (D) Spheroid formation of Fa2N-4 spheroids or Fa2N-4 MCHSs. (E) Expression of EMT-related proteins (E-cadherin, Snail), mesenchymal-related proteins (vimentin, α-SMA), EndMT-related protein (CD31), and ECM-related protein (collagen I) in hepatocyte spheroids and MCHSs. (F) Spheroids treated with 0.5 or 1 µM retinoic acid and forskolin in MCTS (upper panel) and MCHS (bottom panel). All images were obtained using the Operetta CLS system. Data are expressed as means ± SD (n = 3). ^**P < 0.01, and ^***P < 0.001 compared to the control group.