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. 2019 Feb 28;9:2990. doi: 10.1038/s41598-019-39643-y

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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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CRMP2 expression downstream of PTP4A3 reduces cell migration and invasiveness. (A) Western blot showing the knockdown of CRMP2 in OCM-1 cells expressing EGFP, EGFP-PTP4A3, or EGFP-C104S. Twenty micrograms of protein extract was loaded. The detection of α-tubulin was performed as a loading control. (B) The mean overall speed of OCM-1 cells was assessed by a 2D random cell migration assay by time-lapse video microscopy. Data are shown as the mean values +/− SD. ~100 cells were tracked per condition. ***p < 0.001, Student’s t-test. (C) Western blot showing the knockdown of CRMP2 in human PDX-MP41 cells and endogenous expression of PTP4A3. Twenty micrograms of protein extract was loaded. The detection of β-actin was performed as a loading control. (D) The mean overall speed of MP41 cells was assessed by a 2D random cell migration assay by time-lapse video microscopy. Data are shown as the mean values +/− SD. ~100 cells were tracked per condition. ***p < 0.001, Student’s t-test. (E) Quantitative analysis of the relative invasiveness of EGFP-PTP4A3shCtrl and EGFP-PTP4A3shCRMP2 cells. Cells (0.25 × 10⁶) were inoculated into the CAM of chick embryos and the presence of human cells assessed by real-time PCR analysis of chick GAPDH and human alu sequences in the chick femur DNA. Values for the calibrator were arbitrarily defined as 1. The graphs show the means +/− SD. *P < 0.05 (embryos n = 14/cell line), Student test. (F) MMP14 localization on the cell surface assessed by flow cytometry of nonpermeabilized OCM-1 cells. n > 8000 cells analyzed, (****p < 0.0001), Student test.