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. 2020 Feb 10;10:2259. doi: 10.1038/s41598-020-58264-4

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© The Author(s) 2020

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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Subnuclear localization of GLS2. Confocal microscopy of HepG2 cells treated with PMA. (A) GLS2-Nuclear speckles. Double immunofluorescence labeling with anti-GLS2 antibodies (left, green) and anti-nuclear speckles antibodies (center, red); right, view of double labeled cells showing only a few overlapping spots between the two immunolabels. (B) GLS2-Nucleolus. Double immunofluorescence labeling with anti-GLS2 antibodies (left, green) and antibodies against fibrillarin specific for cell nucleoli (center, red); right, merge view of double labeled cells: the nucleolar marker did not overlap with the nuclear GLS2 mark. (C) GLS2-Nucleoplasm. Confocal microscopy of HepG2 cells treated with PMA after double fluorescence labeling with anti-GLS2 antibodies (left, green) and anti-JMJD5 histone demethylase antibodies, a marker specific for cell nucleoplasm (center, red). (D) Double immunofluorescence labeling and confocal microscopy of HepG2 cell employing anti-VAMP8 (green) and anti-GLS2 (red) antibodies: the merge panel shows a high degree of overlapping marks.