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. 2019 Nov 29;9:17889. doi: 10.1038/s41598-019-54478-3

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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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NCAPH-knockdown cells fail to progress of mature chromosome condensation (MCC) from poorly condensed chromosomes. (A) General outline of the procedure followed for thymidine double block synchronization of MIA PaCa-2 cells and their cell cycle analysis. Cells were transfected with the indicated siRNAs, and after release from the second thymidine block, cells were assayed at different time points ranging from 0 to 8 h. (B) Cells were harvested for PI staining and analyzed by flow cytometry to determine the cell cycle fraction at 0, 4, and 8 h after release. Flow cytometry plots and data are representative of at least three separate experiments. (C) The percentages of cells in G₁, S, and G₂/M phases are indicated. (D,E) Frequencies of poorly condensed chromosomes and mature condensed chromosomes cells in control and NCAPH-knockdown cells. Cells were transfected with the indicated siRNA, treated with one thymidine block (36 h), released for 3 h and then harvested. For accurate quantification, more than 200 cells captured in at least three different fields were analyzed. Values represent means ± SEMs. *P < 0.005, ***P < 0.0001. Statistical analysis was performed using one-way analysis of variance followed by Bonferroni’s multiple comparison tests.