Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

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

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 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/.

PMC Copyright notice

Calpain-6 promotes angiogenesis in vitro. (a,b) Secreted VEGF by calpain-6 increased HUVEC migration and invasion. HUVECs seeded on Transwells for the migration assays (a) or on Matrigel-coated Transwells for the invasion assays (b) were co-cultured with stable calpain-6-overexpressing or control vector HEK293 cells for 3 h. The number of migrated or invaded cells was photographed under a light microscope, and mean values were determined. (c) Calpain-6 overexpression in HUVECs promotes tube formation in vitro. Tube formation assays were performed with HUVECs transfected with control vector or calpain-6 expressing vector. As a positive control, HUVECs were incubated with recombinant human VEGF (10 ng/mL). The formation of tubular structures was captured by an inverted light microscope, and tube lengths were quantified by Inform. (d) Calpain-6 expression increased VEGF secretion in HUVECs transected with a calpain-6 expressing vector. VEGF protein in the conditioned medium of HUVECs transfected with control vector or calpain-6 expressing vector was analyzed by ELISA and the results were expressed as mean VEGF produced in the conditioned medium. Data represent the mean ± SD from three independent experiments. (e) Silencing of calpain-6 expression by siRNA. Calpain-6 stable HEK 293 cells were transfected with scrambled (SCR) and calpain-6 siRNA, respectively, and then analyzed the relative calpain-6 mRNA expression by real-time PCR. The results were expressed as the mean values. (f) Silencing of calpain-6 by siRNA decreased the VEGF secretion in HEK293 cells and leads to reduction in tube formation in HUVECs. The quantitation of VEGF secreted in the CM of stable HEK293-calpain-6 cells by calpain-6 siRNA was analyzed by ELISA and the results are expressed as mean VEGF produced in the conditioned medium. Data represent the mean ± SD of three independent experiments.