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. 2017 Nov 2;7:14932. doi: 10.1038/s41598-017-06500-9

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© The Author(s) 2017

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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GP73 regulates the transcriptional activity of SREBPs and lipogenesis. (a) Immunoblotting analysis of SREBPs activation in HepG2 cells transfected with Flag-GP73 at the indicated doses. α-Tubulin was used as equal loading control. (b,c) SREBP-1 promoter activity in HepG2 (b) or HL7702 (c) cells transfected with Flag-vector or Flag-GP73 under conditions of sterol depletion or repletion. The luciferase activity was measured 36 hrs post transfection. The value was normalized with the corresponding transfection efficiency. (d,f,h) QRT-PCR analysis of HMGR (d), FASN2 (f), and ACC1 (h) mRNA abundance in HepG2 cells transfected with Flag-vector or Flag-GP73 for 24 hrs. (e,g,i) QRT-PCR analysis of HMGR (e), FASN2 (g), and ACC1 (i) mRNA abundance in 293T cells transfected with Flag-vector or Flag-GP73 for 24 hrs. (j) Fluorescence microscopy of Filipin staining in HepG2 cells transfected with Flag-GP73. Cells were collected at indicated hrs post transfection. (k,l) Amplex Red cholesterol assay of cellular cholesterol concentrations in HepG2 (k) or HL7702 (l) cells transfected with Flag-vector or Flag-GP73. Cells were collected at indicated hrs post transfection. Values were normalized to total cell proteins from control cells transfected with Flag-vector. Cell-based studies were performed at least three independent times with comparable results. Data represent mean ± SEM. Student’s t test was used for statistical analysis: **p < 0.01.