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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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Effects of NCAPH on colony formation ability and proliferation of PC cells. (A) MIA PaCa-2 cells were transfected with three different siRNA constructs to knockdown NCAPH expression, and the expression levels of NCAPH protein were measured 48 h later by western blotting. Cell lysates were immunoblotted with the indicated antibodies. (B) Colony formation assays were performed on MIA PaCa-2 and PANC-1 cells transfected with the control and three different NCAPH siRNAs (NCAPH siRNA_1, siRNA_2, and siRNA _3). (C) Colony formation assays were performed in AsPC-1, Capan-1, and Capan-2 cells with control siRNA and NCAPH siRNA_3. The colonies were imaged and counted. Values represent means ± SEMs. Statistical analysis was performed using unpaired t-tests. ***P < 0.0001. (D) MTT assays were performed to determine proliferation of control and NCAPH-knockdown PC cells for 5 or 7 days. The curve was based on biological triplicates with values expressed as means ± SEMs. *P < 0.005, ***P < 0.0001. Statistical analysis was performed using two-way analysis of variance with Bonferroni’s multiple comparison tests.