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. 2019 Apr 5;10(4):312. doi: 10.1038/s41419-019-1541-1

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© The Author(s) 2019

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. 5 — a 97H cells were infected with adenovirus-expressing GFP (Ad-GFP) or SET8 tagged with Flag epitope (Ad-SET8). Cells were harvested, and nuclear extracts were sequentially immunoprecipitated with Anti-FLAG (M2 agarose). The SET8-associated proteins were detected by SDS/PAGE and silver staining. b HEK293T cells were ectopically expressed with the indicated plasmids (Flag-SET8 and HA-KLF4). Extracts were immunoprecipitated with the antibody against Flag tag (IP: Flag-SET8/IB: KLF4) or KLF4 (IP: HA-KLF4/IB: Flag-SET8). c Validation of SET8–KLF4 interaction in vivo. Endogenous interaction between SET8 and KLF4 was observed in HCC cells measured by immunoprecipitation. d Colocalization of SET8 and KLF4 in HCC cells detected by confocal microscopy. e mRNA level of KLF4 detected in SET8-overexpressing cells. f, g Analysis of extracellular acidification rate (ECAR) and oxygen consumption rate (OCR) in M3 (left) and 97H (right) cells with either KLF4 overexpression or control vector. h, i The expression levels of genes involved in glucose metabolism in the indicated HCC cell lines with either KLF4 overexpression or control vector detected by real-time PCR and western blot analysis. Data are shown as mean ± SD of three independent experiments. ^*P < 0.05