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. 2018 Jan 8;8:93. doi: 10.1038/s41598-017-18551-z

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© The Author(s) 2017

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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Knockdown of Sema4B reduces colony formation and cell invasion. (A,B) Colony formation assay experiments (n = 3) were performed for sh-cont and two shRNA targeting Sema4B. Forty-eight (48) h after treatment with the shRNA, 400 cells were seeded in each 6-well plate and stained with Giemsa after 18 days. (A) Example of the results after Giemsa staining. (B) Colonies were counted for each well; data represent the median of three experiments (the p values were calculated with the one-tailed Fisher’s Exact Test, n = 400; p < 3.148E-12). (C,D) Invasion assays using Boyden chamber transwell experiments were performed for sh-cont and two shRNAs targeting Sema4B (n = 3). (C) Invading cells were observed after 7 hours. (D) Six pictures were taken of each treatment and invaded cells were counted. Data represent the median number of counted cells relative to the control, with range (n = 6; p < 0.004). Data are presented as medians with range. The p values were calculated with the two-tailed Mann–Whitney Test.