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. 2019 Oct 31;9:15771. doi: 10.1038/s41598-019-52364-6

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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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VEGFA secretion is enhanced by calpain-6 expression. (a) Calpain-6 increased VEGF secretion in a dose-dependent manner. HEK 293 cells were transiently transfected with calpain-6-overexpressing and control vector and then calpain-6 protein levels were confirmed by immunoblot analysis and ELISA in the cell lysates. (b) Establishment of vector stable control vector and calpain-6 overexpressing HEK293 cells. Calpain-6 overexpressing and control vector only expressing HEK293 cells were further selected in the presence of G418-containing culture medium for 3 weeks and then monoclonal vector control and calpain-6 expressing monoclonal cells were expanded. Conditioned medium (CM) was harvested and then processed for VEGF quantitation using an ELISA analysis. Data represent the mean ± SD of three independent experiments.