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The fibronectin ED-A domain enhances recruitment of latent TGF-β-binding protein-1 to the fibroblast matrix.

J. Cell. Sci.. 2018; 
Klingberg Franco,Chau Grace,Walraven Marielle,Boo Stellar,Koehler Anne,Chow Melissa L,Olsen Abby L,Im Michelle,Lodyga Monika,Wells Rebecca G,White Eric S,Hinz B
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PCR Cloning and Subcloning Primary antibodies used in this study were directed against: α-SMA (clone SM1, a kind gift of Giulio Gabbiani, University of Geneva, Switzerland), (total) FN (rbAb, Sigma-Aldrich, F3648, 1:100), ED-A FN (mAb, clone IST-9, Santa Cruz Biotech, Dallas, TX, sc-59826, 1:100), ED-B FN (mAb, clone BC-1, Abcam, Cambridge, MA, ab154210, 1:100), FN HepII (mIgG1, clone Ab32, Thermo Fisher CSI 005-32-02) LTBP-1 (mAb, R&D Systems, Minneapolis, MN, MAB388, 1:100; and rbAb39, a very generous gift from Carl-Hendrik Heldin, Uppsala University, Sweden, 1:200), HIS (mAb A00186 and rbAb A00174, Genscript, Piscataway, NJ, 1:200), fibrillin-1 (rF6H ID 157, a kind gift from D. Reinhardt, McGill University, Montreal, Canada, 1:50), and vimentin (mAb, Dako, Burlington, ON, M0725, 1:400). Get A Quote

摘要

Dysregulated secretion and extracellular activation of TGF-β1 stimulates myofibroblasts to accumulate disordered and stiff extracellular matrix (ECM) leading to fibrosis. Fibronectin immobilizes latent TGF-β-binding protein-1 (LTBP-1) and thus stores TGF-β1 in the ECM. Because the ED-A fibronectin splice variant is prominently expressed during fibrosis and supports myofibroblast activation, we investigated whether ED-A promotes LTBP-1-fibronectin interactions. Using stiffness-tuneable substrates for human dermal fibroblast cultures, we showed that high ECM stiffness promotes expression and colocalization of LTBP-1 and ED-A-containing fibronectin. When rescuing fibronectin-depleted fibroblasts with specif... More

关键词

Fibrosis,Growth factor activation,Myofibroblast,TGF-β1,Transforming growth factor β1,Wound hea