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. 2018 Oct 25;29(1):8–22. doi: 10.1038/s41422-018-0103-x

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© The Author(s) 2018

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — SIRT1 is essential for hepatocyte-to-LPC conversion and expansion. a Immunofluorescence images of SIRT1 and cleaved-caspase-3 at day 4 in cells transfected with control siRNA (siCTL) or siRNA-targeting SIRT1 (siSIRT1). Scale bars, 20 μm. b Western blot analysis of SIRT1 and cleaved-caspase-3 expression as in (a). c QPCR analyses for the expression of P21, FOXO3 and NANOG in cells transfected with siCTL or siSIRT1. Error bars represent s.d.; n = 3 donors (two-tailed unpaired t-test, ***P < 0.001). d CCK-8 analyses demonstrating suppression of cell proliferation in the presence of siSIRT1. Error bars represent s.d.; n = 5 technical replicates from one donor. e Light microscopy images show a reduction in clone formation following inhibition of SIRT1 by crystal violet staining. Scale bars, 10 mm. f Schematic of the 7 factors (7 F) withdrawal assay during passage. g CCK-8 analyses demonstrating suppression of cell proliferation in the absence of the 7 F within 48 h. Error bars represent s.d.; n = 5 technical replicates from one donor. h Light microscopy shows reduced cell proliferation in response to the withdrawal of the 7 F at 24 h and 48 h. Scale bars, 100 µm. i Immunofluorescence images of SIRT1 and cleaved-caspase-3 in HepLPCs cultured in TEM or TEM-7F at 24 h. Scale bars, 20μm. j Western blot analysis of SIRT1 and cleaved caspase-3 expression as in (i). k QPCR analyses for the expression of P21, FOXO3 and NANOG as in (i). Error bars represent s.d.; n = 3 donors (two-tailed unpaired t-test, **P < 0.01, ***P < 0.001)