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. 2018 Aug 17;8:12337. doi: 10.1038/s41598-018-30886-9

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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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Nuclear function of novex-3 as a cell cycle promoter is ascribed to that in interphase, but not during mitosis. (a) A representative time-lapse recording of cell division dynamics of the E16 fCM doubly expressing novex-3-Nterm-GFP and sarcomeric α-actinin-mCherry (as a CM marker). In contrast to the clear nuclear expression observed during interphase (arrow), novex-3-Nterm expression was not localised to specific structures (spindles, spindle matrix, centrioles, or chromosomes) during mitosis; instead, it was diffusely spread throughout the cytoplasm following nuclear envelope breakdown at prometaphase. At telophase, when the nuclear envelope reassembled, the novex-3-Nterm re-accumulated in the newly formed daughter nuclei (arrows in telophase and cytokinesis), where it was maintained over the next interphase (arrows). The number on each panel indicates the time in (hours: minutes) elapsed from the time indicated in the first panel. Scale bar = 50 µm. (b) Cultured fCMs (E17–E18) were triple-stained with novex-3, sarcomeric α-actinin (as a CM marker), and DAPI. DAPI staining clearly defined each phase in mitosis, including prophase, metaphase, anaphase, and subsequent cytokinesis. In interphase fCMs, novex-3 was expressed both in nuclei and sarcomere (arrows). At this time point, sarcomere structure was intact, as indicated by α-actinin staining. In prophase, when chromosome condensation was visible as dot-like puncta by DAPI staining (arrow), novex-3 was still expressed in fCM nuclei (arrow), while sarcomere structure has begun to disassemble. After the nuclear envelope breakdown at prometaphase, novex-3 did not localise to the specific structures such as spindles, spindle matrix, centrioles, or chromosomes; instead it diffused throughout the cytoplasm. When the nuclear envelope reassembled during cytokinesis, novex-3 re-accumulated in the newly formed daughter nuclei (arrows). Scale bar = 20 µm.