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. 2018 Aug 24;8:12780. doi: 10.1038/s41598-018-29448-w

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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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PIASγ reduces the cytosolic HNF-1A protein level in a dose- dependent manner. (a) Effect of PIASγ on HNF-1A protein in cleared lysates from transiently transfected HEK293 cells, with V5-tagged HNF-1A together with increasing amounts of Flag-tagged PIASγ (0.5–2 µg), and analyzed by SDS-PAGE and immunoblotting using indicated antibodies. Full-length blots are presented in Supplementary Fig. S9. (b) The level of HNF-1A was normalized to actin and presented relative to the level of HNF-1A alone. The results are shown as mean of four independent experiments ±SD (n = 4). *Indicates p < 0.05. (c) HEK293 cells were transiently transfected with V5-tagged HNF-1A in the presence or absence of HA-SUMO-3 and Flag-tagged PIASγ. Post-transfection, cells were treated with 10 µg MG132 or DMSO for 8 hours, and protein from cleared lysates was analyzed by SDS-PAGE and immunoblotting using indicated antibodies. Full-length blots are presented in Supplementary Fig. S10. (d) Quantification of proteins shown in c) by densiometric analysis. The level of HNF-1A was normalized to the loading control (actin). Each column represents the mean fold difference in the MG132-treated samples versus DMSO-treated control on two different experimental days ± SD (n = 2).