Prestress in the extracellular matrix sensitizes latent TGF-β1 for activation

Journal of Cell Biology

Volumn 207, Issue 2, 2014, Pages 283-297

  Klingberg, Franco ^a   Chow, Melissa L ^a   Koehler, Anne ^a   Boo, Stellar ^a   Buscemi, Lara ^b   Quinn, Thomas M ^c   Costell, Mercedes ^d   Alman, Benjamin A ^e   Genot, Elisabeth ^f   Hinz, Boris ^a  

^a UNIVERSITY OF TORONTO   (Canada)

^b UNIVERSITY OF LAUSANNE   (Switzerland)

^c MCGILL UNIVERSITY   (Canada)

^d UNIVERSITY OF VALENCIA   (Spain)

^e HOSPITAL FOR SICK CHILDREN   (Canada)

^f UNIVERSITÉ DE BORDEAUX   (France)

Author keywords

[No Author keywords available]

Indexed keywords

ACTIN; INTEGRIN; MYOSIN; TRANSFORMING GROWTH FACTOR BETA1;

ANIMAL EXPERIMENT; ARTICLE; BIOAVAILABILITY; CELL ADHESION; CELL CULTURE; CELL DIFFERENTIATION; COMPARATIVE STUDY; CONFORMATIONAL TRANSITION; CONTROLLED STUDY; EXTRACELLULAR MATRIX; FEMALE; HUMAN; HUMAN TISSUE; IMMUNOFLUORESCENCE; IMMUNOPRECIPITATION; MECHANICAL STRESS; MYOFIBROBLAST; NONHUMAN; PROTEIN PURIFICATION; RAT; STRESS FIBER; WESTERN BLOTTING; ANIMAL; CYTOLOGY; HEK293 CELL LINE; MECHANOTRANSDUCTION; METABOLISM; PHYSIOLOGY; WISTAR RAT;

ANIMALS; CELL DIFFERENTIATION; CELLS, CULTURED; EXTRACELLULAR MATRIX; HEK293 CELLS; HUMANS; INTEGRINS; MECHANOTRANSDUCTION, CELLULAR; MYOFIBROBLASTS; RATS, WISTAR; TRANSFORMING GROWTH FACTOR BETA1;

EID: 84910601354     PISSN: 00219525     EISSN: 15408140     Source Type: Journal    
DOI: 10.1083/jcb.201402006     Document Type: Article

References (74)

1. Abe, M., J.G. Harpel, C.N. Metz, I. Nunes, D.J. Loskutoff, and D.B. Rifkin. 1994. An assay for transforming growth factor-β using cells transfected with a plasminogen activator inhibitor-1 promoter-luciferase construct. Anal. Biochem. 216:276-284. http://dx.doi.org/10.1006/abio.1994.1042
2. 6-mediated activation of latent TGF-β requires the latent TGF-β binding protein-1. J. Cell Biol. 165:723-734. http://dx.doi.org/10.1083/jcb.200312172
3. Baldwin, A.K., A. Simpson, R. Steer, S.A. Cain, and C.M. Kielty. 2013. Elastic fibres in health and disease. Expert Rev. Mol. Med. 15:e8. http://dx.doi.org/10.1017/erm.2013.9
4. Balestrini, J.L., S. Chaudhry, V. Sarrazy, A. Koehler, and B. Hinz. 2012. The mechanical memory of lung myofibroblasts. Integr. Biol. (Camb.). 4:410-421. http://dx.doi.org/10.1039/c2ib00149g
5. Berry, M.F., A.J. Engler, Y.J. Woo, T.J. Pirolli, L.T. Bish, V. Jayasankar, K.J. Morine, T.J. Gardner, D.E. Discher, and H.L. Sweeney. 2006. Mesenchymal stem cell injection after myocardial infarction improves myocardial compliance. Am. J. Physiol. Heart Circ. Physiol. 290:H2196- H2203. http://dx.doi.org/10.1152/ajpheart.01017.2005
6. Booth, A.J., R. Hadley, A.M. Cornett, A.A. Dreffs, S.A. Matthes, J.L. Tsui, K. Weiss, J.C. Horowitz, V.F. Fiore, T.H. Barker, et al. 2012. Acellular normal and fibrotic human lung matrices as a culture system for in vitro investigation. Am. J. Respir. Crit. Care Med. 186:866-876. http://dx.doi.org/10.1164/rccm.201204-0754OC
7. Brunner, G., and R. Blakytny. 2004. Extracellular regulation of TGF-β activity in wound repair: growth factor latency as a sensor mechanism for injury. Thromb. Haemost. 92:253-261.
8. Buscemi, L., D. Ramonet, F. Klingberg, A. Formey, J. Smith-Clerc, J.J. Meister, and B. Hinz. 2011. The single-molecule mechanics of the latent TGF-β1 complex. Curr. Biol. 21:2046-2054. http://dx.doi.org/10.1016/j.cub.2011.11.037
9. Chaudhry, S.S., S.A. Cain, A. Morgan, S.L. Dallas, C.A. Shuttleworth, and C.M. Kielty. 2007. Fibrillin-1 regulates the bioavailability of TGFβ1. J. Cell Biol. 176:355-367. http://dx.doi.org/10.1083/jcb.200608167
10. Chen, Q., P. Sivakumar, C. Barley, D.M. Peters, R.R. Gomes, M.C. Farach-Carson, and S.L. Dallas. 2007. Potential role for heparan sulfate proteoglycans in regulation of transforming growth factor-beta (TGF-β) by modulating assembly of latent TGF-β-binding protein-1. J. Biol. Chem. 282:26418-26430. http://dx.doi.org/10.1074/jbc.M703341200
11. Dallas, S.L., D.R. Keene, S.P. Bruder, J. Saharinen, L.Y. Sakai, G.R. Mundy, and L.F. Bonewald. 2000. Role of the latent transforming growth factor β binding protein 1 in fibrillin-containing microfibrils in bone cells in vitro and in vivo. J. Bone Miner. Res. 15:68-81. http://dx.doi.org/10.1359/jbmr.2000.15.1.68
12. Dallas, S.L., P. Sivakumar, C.J. Jones, Q. Chen, D.M. Peters, D.F. Mosher, M.J. Humphries, and C.M. Kielty. 2005. Fibronectin regulates latent transforming growth factor-beta (TGF β) by controlling matrix assembly of latent TGF β-binding protein-1. J. Biol. Chem. 280:18871-18880. http://dx.doi.org/10.1074/jbc.M410762200
13. Discher, D., C. Dong, J.J. Fredberg, F. Guilak, D. Ingber, P. Janmey, R.D. Kamm, G.W. Schmid-Schonbein, and S. Weinbaum. 2009. Biomechanics: cell research and applications for the next decade. Ann. Biomed. Eng. 37:847- 859. http://dx.doi.org/10.1007/s10439-009-9661-x
14. Doyle, J.J., E.E. Gerber, and H.C. Dietz. 2012. Matrix-dependent perturbation of TGFβ signaling and disease. FEBS Lett. 586:2003-2015. http://dx.doi.org/10.1016/j.febslet.2012.05.027
15. Fassler, R., and M. Meyer. 1995. Consequences of lack of beta 1 integrin gene expression in mice. Genes Dev. 9:1896-1908. http://dx.doi.org/10.1101/gad.9.15.1896
16. Fassler, R., M. Pfaff, J. Murphy, A.A. Noegel, S. Johansson, R. Timpl, and R. Albrecht. 1995. Lack of β1 integrin gene in embryonic stem cells affects morphology, adhesion, and migration but not integration into the inner cell mass of blastocysts. J. Cell Biol. 128:979-988. http://dx.doi.org/10.1083/jcb.128.5.979
17. Follonier Castella, L.F., L. Buscemi, C. Godbout, J.J. Meister, and B. Hinz. 2010a. A new lock-step mechanism of matrix remodelling based on subcellular contractile events. J. Cell Sci. 123:1751-1760. http://dx.doi.org/10.1242/jcs.066795
18. Follonier Castella, L., G. Gabbiani, C.A. McCulloch, and B. Hinz. 2010b. Regulation of myofibroblast activities: calcium pulls some strings behind the scene. Exp. Cell Res. 316:2390-2401. http://dx.doi.org/10.1016/j.yexcr.2010.04.033
19. 6-mediated activation of latent TGF-β complexes containing LTBP-1. FASEB J. 19:1798- 1808. http://dx.doi.org/10.1096/fj.05-4134com
20. Gerber, E.E., E.M. Gallo, S.C. Fontana, E.C. Davis, F.M. Wigley, D.L. Huso, and H.C. Dietz. 2013. Integrin-modulating therapy prevents fibrosis and autoimmunity in mouse models of scleroderma. Nature. 503:126-130. http://dx.doi.org/10.1038/nature12614
21. Giacomini, M.M., M.A. Travis, M. Kudo, and D. Sheppard. 2012. Epithelial cells utilize cortical actin/myosin to activate latent TGF-β through integrin α(v)β(6)-dependent physical force. Exp. Cell Res. 318:716-722. http://dx.doi.org/10.1016/j.yexcr.2012.01.020
22. Godbout, C., L. Follonier Castella, E.A. Smith, N. Talele, M.L. Chow, A. Garonna, and B. Hinz. 2013. The mechanical environment modulates intracellular calcium oscillation activities of myofibroblasts. PLoS ONE. 8:e64560. http://dx.doi.org/10.1371/journal.pone.0064560
23. Godyna, S., D.M. Mann, and W.S. Argraves. 1995. A quantitative analysis of the incorporation of fibulin-1 into extracellular matrix indicates that fibronectin assembly is required. Matrix Biol. 14:467-477. http://dx.doi.org/10.1016/0945-053X(95)90004-7
24. Goffin, J.M., P. Pittet, G. Csucs, J.W. Lussi, J.J. Meister, and B. Hinz. 2006. Focal adhesion size controls tension-dependent recruitment of β-smooth muscle actin to stress fibers. J. Cell Biol. 172:259-268. http://dx.doi.org/10.1083/jcb.200506179
25. Henderson, N.C., and D. Sheppard. 2013. Integrin-mediated regulation of TGFβ in fibrosis. Biochim. Biophys. Acta. 1832:891-896. http://dx.doi.org/10.1016/j.bbadis.2012.10.005
26. Henderson, N.C., T.D. Arnold, Y. Katamura, M.M. Giacomini, J.D. Rodriguez, J.H. McCarty, A. Pellicoro, E. Raschperger, C. Betsholtz, P.G. Ruminski, et al. 2013. Targeting of βv integrin identifies a core molecular pathway that regulates fibrosis in several organs. Nat. Med. 19:1617-1624. http://dx.doi.org/10.1038/nm.3282
27. Hinz, B. 2012. Mechanical aspects of lung fibrosis: a spotlight on the myofibroblast. Proc. Am. Thorac. Soc. 9:137-147. http://dx.doi.org/10.1513/pats.201202-017AW
28. Hinz, B. 2013. It has to be the αv: myofibroblast integrins activate latent TGF-β1. Nat. Med. 19:1567-1568. http://dx.doi.org/10.1038/nm.3421
29. Hinz, B., G. Celetta, J.J. Tomasek, G. Gabbiani, and C. Chaponnier. 2001a. α-smooth muscle actin expression upregulates fibroblast contractile activity. Mol. Biol. Cell. 12:2730-2741. http://dx.doi.org/10.1091/mbc.12.9.2730
30. Hinz, B., D. Mastrangelo, C.E. Iselin, C. Chaponnier, and G. Gabbiani. 2001b. Mechanical tension controls granulation tissue contractile activity and myofibroblast differentiation. Am. J. Pathol. 159:1009-1020. http://dx.doi.org/10.1016/S0002-9440(10)61776-2
31. Hinz, B., S.H. Phan, V.J. Thannickal, A. Galli, M.L. Bochaton-Piallat, and G. Gabbiani. 2007. The myofibroblast: one function, multiple origins. Am. J. Pathol. 170:1807-1816. http://dx.doi.org/10.2353/ajpath.2007.070112
32. Hinz, B., S.H. Phan, V.J. Thannickal, M. Prunotto, A. Desmouliere, J. Varga, O. De Wever, M. Mareel, and G. Gabbiani. 2012. Recent developments in myofibroblast biology: paradigms for connective tissue remodeling. Am. J. Pathol. 180:1340-1355. http://dx.doi.org/10.1016/j.ajpath.2012.02.004
33. Hyytiäinen, M., C. Penttinen, and J. Keski-Oja. 2004. Latent TGF-β binding proteins: extracellular matrix association and roles in TGF-β activation. Crit. Rev. Clin. Lab. Sci. 41:233-264. http://dx.doi.org/10.1080/10408360490460933
34. Ilić, D., B. Kovacic, K. Johkura, D.D. Schlaepfer, N. Tomasević, Q. Han, J.B. Kim, K. Howerton, C. Baumbusch, N. Ogiwara, et al. 2004. FAK promotes organization of fibronectin matrix and fibrillar adhesions. J. Cell Sci. 117:177-187. http://dx.doi.org/10.1242/jcs.00845
35. Isogai, Z., R.N. Ono, S. Ushiro, D.R. Keene, Y. Chen, R. Mazzieri, N.L. Charbonneau, D.P. Reinhardt, D.B. Rifkin, and L.Y. Sakai. 2003. Latent transforming growth factor β-binding protein 1 interacts with fibrillin and is a microfibril-associated protein. J. Biol. Chem. 278:2750-2757. http://dx.doi.org/10.1074/jbc.M209256200
36. Jenkins, G. 2008. The role of proteases in transforming growth factor-β activation. Int. J. Biochem. Cell Biol. 40:1068-1078. http://dx.doi.org/10.1016/j.biocel.2007.11.026
37. Judge, D.P., N.J. Biery, D.R. Keene, J. Geubtner, L. Myers, D.L. Huso, L.Y. Sakai, and H.C. Dietz. 2004. Evidence for a critical contribution of haploinsufficiency in the complex pathogenesis of Marfan syndrome. J. Clin. Invest. 114:172-181. http://dx.doi.org/10.1172/JCI200420641
38. Kiema, T., Y. Lad, P. Jiang, C.L. Oxley, M. Baldassarre, K.L. Wegener, I.D. Campbell, J. Ylanne, and D.A. Calderwood. 2006. The molecular basis of filamin binding to integrins and competition with talin. Mol. Cell. 21:337-347. http://dx.doi.org/10.1016/j.molcel.2006.01.011
39. Kim, H., A. Sengupta, M. Glogauer, and C.A. McCulloch. 2008. Filamin A regulates cell spreading and survival via β1 integrins. Exp. Cell Res. 314:834- 846. http://dx.doi.org/10.1016/j.yexcr.2007.11.022
40. Klingberg, F., B. Hinz, and E.S. White. 2013. The myofibroblast matrix: implications for tissue repair and fibrosis. J. Pathol. 229:298-309. http://dx.doi.org/10.1002/path.4104
41. Koli, K., M. Hyytiainen, M.J. Ryynanen, and J. Keski-Oja. 2005. Sequential deposition of latent TGF-β binding proteins (LTBPs) during formation of the extracellular matrix in human lung fibroblasts. Exp. Cell Res. 310:370-382. http://dx.doi.org/10.1016/j.yexcr.2005.08.008
42. Maeda, T., T. Sakabe, A. Sunaga, K. Sakai, A.L. Rivera, D.R. Keene, T. Sasaki, E. Stavnezer, J. Iannotti, R. Schweitzer, et al. 2011. Conversion of mechanical force into TGF-β-mediated biochemical signals. Curr. Biol. 21:933-941. http://dx.doi.org/10.1016/j.cub.2011.04.007
43. Majd, H., T.M. Quinn, P.J. Wipff, and B. Hinz. 2011. Dynamic expansion culture for mesenchymal stem cells. Methods Mol. Biol. 698:175-188. http://dx.doi.org/10.1007/978-1-60761-999-4_14
44. Massam-Wu, T., M. Chiu, R. Choudhury, S.S. Chaudhry, A.K. Baldwin, A. McGovern, C. Baldock, C.A. Shuttleworth, and C.M. Kielty. 2010. Assembly of fibrillin microfibrils governs extracellular deposition of latent TGF β. J. Cell Sci. 123:3006-3018. http://dx.doi.org/10.1242/jcs.073437
45. Nishimura, S.L. 2009. Integrin-mediated transforming growth factor-β activation, a potential therapeutic target in fibrogenic disorders. Am. J. Pathol. 175:1362-1370. http://dx.doi.org/10.2353/ajpath.2009.090393
46. Ono, R.N., G. Sengle, N.L. Charbonneau, V. Carlberg, H.P. Bachinger, T. Sasaki, S. Lee-Arteaga, L. Zilberberg, D.B. Rifkin, F. Ramirez, et al. 2009. Latent transforming growth factor β-binding proteins and fibulins compete for fibrillin-1 and exhibit exquisite specificities in binding sites. J. Biol. Chem. 284:16872-16881. http://dx.doi.org/10.1074/jbc.M809348200
47. Rajshankar, D., G.P. Downey, and C.A. McCulloch. 2012. IL-1β enhances cell adhesion to degraded fibronectin. FASEB J. 26:4429-4444. http://dx.doi.org/10.1096/fj.12-207381
48. Ramirez, F., and D.B. Rifkin. 2009. Extracellular microfibrils: contextual platforms for TGFβ and BMP signaling. Curr. Opin. Cell Biol. 21:616-622. http://dx.doi.org/10.1016/j.ceb.2009.05.005
49. Robertson, I.B., and D.B. Rifkin. 2013. Unchaining the beast; insights from structural and evolutionary studies on TGFβ secretion, sequestration, and activation. Cytokine Growth Factor Rev. 24:355-372. http://dx.doi.org/10.1016/j.cytogfr.2013.06.003
50. Rosenzweig, D.H., M. Matmati, G. Khayat, S. Chaudhry, B. Hinz, and T.M. Quinn. 2012. Culture of primary bovine chondrocytes on a continuously expanding surface inhibits dedifferentiation. Tissue Eng. Part A. 18:2466-2476. http://dx.doi.org/10.1089/ten.tea.2012.0215
51. Sabatier, L., D. Chen, C. Fagotto-Kaufmann, D. Hubmacher, M.D. McKee, D.S. Annis, D.F. Mosher, and D.P. Reinhardt. 2009. Fibrillin assembly requires fibronectin. Mol. Biol. Cell. 20:846-858. http://dx.doi.org/10.1091/mbc.E08-08-0830
52. Sabatier, L., J. Djokic, C. Fagotto-Kaufmann, M. Chen, D.S. Annis, D.F. Mosher, and D.P. Reinhardt. 2013. Complex contributions of fibronectin to initiation and maturation of microfibrils. Biochem. J. 456:283-295. http://dx.doi.org/10.1042/BJ20130699
53. Saharinen, J., M. Hyytiainen, J. Taipale, and J. Keski-Oja. 1999. Latent transforming growth factor-β binding proteins (LTBPs)-structural extracellular matrix proteins for targeting TGF-β action. Cytokine Growth Factor Rev. 10:99-117. http://dx.doi.org/10.1016/S1359-6101(99)00010-6
54. Sakai, T., K.J. Johnson, M. Murozono, K. Sakai, M.A. Magnuson, T. Wieloch, T. Cronberg, A. Isshiki, H.P. Erickson, and R. Fassler. 2001. Plasma fibronectin supports neuronal survival and reduces brain injury following transient focal cerebral ischemia but is not essential for skinwound healing and hemostasis. Nat. Med. 7:324-330. http://dx.doi.org/10.1038/85471
55. Sarrazy, V., A. Koehler, M.L. Chow, E. Zimina, C.X. Li, H. Kato, C.A. Caldarone, and B. Hinz. 2014. Integrins αvβ5 and αvβ3 promote latent TGF-β1 activation by human cardiac fibroblast contraction. Cardiovasc. Res. 102:407-417. http://dx.doi.org/10.1093/cvr/cvu053
56. Shi, M., J. Zhu, R. Wang, X. Chen, L. Mi, T. Walz, and T.A. Springer. 2011. Latent TGF-β structure and activation. Nature. 474:343-349. http://dx.doi.org/10.1038/nature10152
57. Shifrin, Y., P.D. Arora, Y. Ohta, D.A. Calderwood, and C.A. McCulloch. 2009. The role of FilGAP-filamin A interactions in mechanoprotection. Mol. Biol. Cell. 20:1269-1279. http://dx.doi.org/10.1091/mbc.E08-08-0872
58. Shimbori, C., J. Gauldie, and M. Kolb. 2013. Extracellular matrix microenvironment contributes actively to pulmonary fibrosis. Curr. Opin. Pulm. Med. 19:446-452. http://dx.doi.org/10.1097/MCP.0b013e328363f4de
59. Singh, P., C. Carraher, and J.E. Schwarzbauer. 2010. Assembly of fibronectin extracellular matrix. Annu. Rev. Cell Dev. Biol. 26:397-419. http://dx.doi.org/10.1146/annurev-cellbio-100109-104020
60. Smith, M.L., D. Gourdon, W.C. Little, K.E. Kubow, R.A. Eguiluz, S. Luna-Morris, and V. Vogel. 2007. Force-induced unfolding of fibronectin in the extracellular matrix of living cells. PLoS Biol. 5:e268. http://dx.doi.org/10.1371/journal.pbio.0050268
61. Storm, C., J.J. Pastore, F.C. MacKintosh, T.C. Lubensky, and P.A. Janmey. 2005. Nonlinear elasticity in biological gels. Nature. 435:191-194. http://dx.doi.org/10.1038/nature03521
62. Taipale, J., K. Miyazono, C.H. Heldin, and J. Keski-Oja. 1994. Latent transforming growth factor-β1 associates to fibroblast extracellular matrix via latent TGF-β binding protein. J. Cell Biol. 124:171-181. http://dx.doi.org/10.1083/jcb.124.1.171
63. Taipale, J., J. Saharinen, K. Hedman, and J. Keski-Oja. 1996. Latent transforming growth factor-beta 1 and its binding protein are components of extracellular matrix microfibrils. J. Histochem. Cytochem. 44:875-889. http://dx.doi.org/10.1177/44.8.8756760
64. Todorovic, V., and D.B. Rifkin. 2012. LTBPs, more than just an escort service. J. Cell. Biochem. 113:410-418. http://dx.doi.org/10.1002/jcb.23385
65. Tomasek, J.J., G. Gabbiani, B. Hinz, C. Chaponnier, and R.A. Brown. 2002. Myofibroblasts and mechano-regulation of connective tissue remodelling. Nat. Rev. Mol. Cell Biol. 3:349-363. http://dx.doi.org/10.1038/nrm809
66. Velling, T., J. Risteli, K. Wennerberg, D.F. Mosher, and S. Johansson. 2002. Polymerization of type I and III collagens is dependent on fibronectin and enhanced by integrins α11β1 and α2β1. J. Biol. Chem. 277:37377- 37381. http://dx.doi.org/10.1074/jbc.M206286200
67. Wipff, P.J., and B. Hinz. 2008. Integrins and the activation of latent transforming growth factor β1 - an intimate relationship. Eur. J. Cell Biol. 87:601-615. http://dx.doi.org/10.1016/j.ejcb.2008.01.012
68. Wipff, P.J., D.B. Rifkin, J.J. Meister, and B. Hinz. 2007. Myofibroblast contraction activates latent TGF-β1 from the extracellular matrix. J. Cell Biol. 179:1311-1323. http://dx.doi.org/10.1083/jcb.200704042
69. Wipff, P.J., H. Majd, C. Acharya, L. Buscemi, J.J. Meister, and B. Hinz. 2009. The covalent attachment of adhesion molecules to silicone membranes for cell stretching applications. Biomaterials. 30:1781-1789. http://dx.doi.org/10.1016/j.biomaterials.2008.12.022
70. Worthington, J.J., J.E. Klementowicz, and M.A. Travis. 2011. TGFβ: a sleeping giant awoken by integrins. Trends Biochem. Sci. 36:47-54. http://dx.doi.org/10.1016/j.tibs.2010.08.002
71. Wynn, T.A. 2008. Cellular and molecular mechanisms of fibrosis. J. Pathol. 214:199-210. http://dx.doi.org/10.1002/path.2277
72. Wynn, T.A., and T.R. Ramalingam. 2012. Mechanisms of fibrosis: therapeutic translation for fibrotic disease. Nat. Med. 18:1028-1040. http://dx.doi.org/10.1038/nm.2807
73. Zhong, C., M. Chrzanowska-Wodnicka, J. Brown, A. Shaub, A.M. Belkin, and K. Burridge. 1998. Rho-mediated contractility exposes a cryptic site in fibronectin and induces fibronectin matrix assembly. J. Cell Biol. 141:539- 551. http://dx.doi.org/10.1083/jcb.141.2.539
74. Zilberberg, L., V. Todorovic, B. Dabovic, M. Horiguchi, T. Courousse, L.Y. Sakai, and D.B. Rifkin. 2012. Specificity of latent TGF-β binding protein (LTBP) incorporation into matrix: role of fibrillins and fibronectin. J. Cell. Physiol. 227:3828-3836. http://dx.doi.org/10.1002/jcp.24094