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

. 2011 Jun 27;193(7):1305–1319. doi: 10.1083/jcb.201011143

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

© 2011 Rivera and Brekken

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

PMC Copyright notice

Figure 7. — αV integrin interacts with endoglin and mediates TGF-β activity in primary SPARC^−/− pericytes. (A) Integrin expression profile of primary pericytes. RT-PCR detection of SPARC, αV integrin (itgav), β1 integrin (itgb1), β3 integrin (itgb3), β6 integrin (itgb6), and RPS6 (rps6). (right) Primary SPARC^+/+ and SPARC^−/− pericytes express αV integrins at their surfaces. 500,000 were seeded onto fibronectin-coated dishes and allowed to adhere overnight in 0.75% serum media. Cells were harvested and prepared for FACS analysis using anti–αV integrin IgG (RMV-7) at 20 µg/ml. Control cells were stained with secondary alone. av, avidin; NC, negative control. (B) αV integrins regulate transwell migration of primary pericytes. Pericytes were allowed to migrate in the presence of 20 µg/ml RMV-7 or 20 µg/ml control IgG. Mean values are presented. Error bars represent SEM (*, P <0.05). (C) αV integrins regulate TGF-β activity in primary SPARC^−/− pericytes. Primary SPARC^+/+ or SPARC^−/− pericytes were incubated overnight in 1.5% serum in the presence of 10 µM SB431542, 20 µg/ml RMV-7, 20 µg/ml control IgG, or vehicle alone as indicated. pSMAD2 and tSMAD2 levels were determined by SDS-PAGE and Western blotting. pSMAD2 levels were normalized to tSMAD2 using ImageJ software. (D) αV integrins associate with endoglin in SPARC^−/− pericytes. Endoglin was immunoprecipitated from lysates harvested from cells in suspension (S) or adhered to plastic (P) or fibronectin (F). Complexes were then subjected to SDS-PAGE and probed for αV integrin by Western blotting. (E) SPARC blocks the αV integrin–endoglin interaction. SPARC^−/− pericytes were incubated with either BSA or recombinant SPARC (rSPARC) at the indicated concentrations for 6 h. Endoglin was immunoprecipitated from cell lysates, and complexes were subjected to SDS-PAGE. αV integrin and SPARC levels were determined by Western blotting. Coprecipitating αV integrin was normalized to αV integrin in whole-cell lysates (WCL) using ImageJ software. IP, immunoprecipitation. All experiments were repeated at least twice with the same results. WT, SPARC^+/+; KO, SPARC^−/−.

HHS Vulnerability Disclosure