Collagen type I is the most abundant component of extracellular matrix in the arterial wall. CO2 in DMEM modified to contain 1 g/l d-glucose, l-glutamine, 110 mg/l sodium pyruvate supplemented with 10% fetal bovine serum (FBS; Gemini, Woodland, CA), and antibiotics. Cells between three and seven passages were used for all experiments. The generation of mice with targeted deletion of PKC was described elsewhere (Miyamoto test was used to evaluate the statistical differences between control and treated groups. Differences with p < 0.05 were considered significant. All 1415562-82-1 experiments were repeated at 1415562-82-1 least three times. Error bars on the graphs represent SEM. Values were expressed as mean SEM. Supplementary Material Supplemental Materials: Click here to view. Acknowledgments We acknowledge the technical support of the Hospital of Special Surgery Musculoskeletal Core Center, which is funded by National Institutes of Health Grant AR46121, and thank Henry Pelish at Harvard Medical School (Boston, MA) for the generous gift of secramine A and Drew Roenneburg for technical assistance in histology. This work was supported by National Institutes of Health Grants R01HL081424 (B.L.), R01GM34107 (E.B.), and T32HL083824 (A.Z.). The hybridoma antibody developed by Heinz Furthmayr was obtained from the Developmental Studies Hybridoma 1415562-82-1 Bank developed under the auspices of the National Institute of Child Health and Human Development and maintained by the Department of Biology, University of Iowa (Iowa City, IA). Abbreviations used: Cdc42cell division cycle 42PKCprotein kinase C SMCsmooth muscle cellTGNtrans-Golgi network Footnotes This article was published online ahead of print in MBoC in Press (http://www.molbiolcell.org/cgi/doi/10.1091/mbc.E11-06-0531) on March 28, 2012. REFERENCES Au JS, Puri C, Ihrke G, Kendrick-Jones J, Buss F. Myosin VI is required for sorting of AP-1B-dependent cargo to the basolateral domain in polarized MDCK cells. J Cell Biol. 2007;177:103C114. [PMC free article] [PubMed]Bai X, Margariti A, Hu Y, Sato Y, Zeng L, Ivetic A, Habi O, 1415562-82-1 Mason JC, Wang X, Xu Q. Protein kinase C deficiency accelerates neointimal lesions of mouse injured artery involving delayed reendothelialization and vasohibin-1 accumulation. Arterioscler Thromb Vasc Biol. 2010;30:2467C2474. [PubMed]Bonfanti L, Mironov AA, Jr, Martinez-Menarguez JA, Martella O, Fusella A, Baldassarre M, Buccione R, Geuze HJ, Mironov AA, Luini A. Procollagen traverses the Golgi stack without leaving the lumen of cisternae: evidence for cisternal maturation. Cell. 1998;95:993C1003. [PubMed]Canty EG, Lu Y, Meadows RS, Shaw MK, Holmes DF, Kadler KE. Coalignment of plasma membrane channels and protrusions (fibripositors) specifies the parallelism of tendon. J Cell Biol. 2004;165:553C563. [PMC free article] [PubMed]Canty EG, Starborg T, Lu Y, Humphries SM, Holmes DF, Meadows RS, Huffman A, O’Toole ET, Kadler KE. Actin filaments are required for fibripositor-mediated collagen fibril alignment in tendon. J Biol Chem. 2006;281:38592C38598. [PubMed]Cao H, Weller S, Orth JD, Chen J, Huang B, Chen JL, Stamnes M, McNiven MA. Actin and Arf1-dependent recruitment of a cortactin-dynamin complex to the Golgi regulates post-Golgi transport. Nat Cell Biol. 2005;7:483C492. [PubMed]Carreno S, Engqvist-Goldstein AE, Zhang CX, McDonald KL, Drubin DG. Actin dynamics coupled to clathrin-coated vesicle formation at the JIP-1 trans-Golgi network. J Cell Biol. 2004;165:781C788. [PMC free article] [PubMed]Clowes MM, Lynch CM, Miller AD, Miller DG, Osborne WR, Clowes AW. Long-term biological response of injured rat carotid artery seeded with smooth muscle cells expressing retrovirally introduced human genes. J Clin Invest. 1994;93:644C651. [PMC free article] [PubMed]De Matteis MA, Luini A. Exiting the Golgi complex. Nat Rev Mol Cell Biol. 2008;9:273C284. [PubMed]Egorov MV, et al. Faciogenital dysplasia protein (FGD1) regulates export of cargo proteins from the Golgi complex via Cdc42 activation. Mol Biol Cell. 2009;20:2413C2427. [PMC free article] [PubMed]Erickson JW, Zhang C, Kahn RA, Evans T, Cerione RA. Mammalian Cdc42 is a brefeldin A-sensitive component of the Golgi apparatus. J Biol Chem. 1996;271:26850C26854. [PubMed]Fukumoto S, Nishizawa Y, Hosoi M, Koyama H, Yamakawa K, Ohno S, Morii H. Protein kinase C delta inhibits the proliferation of vascular smooth muscle cells by suppressing G1 cyclin expression. J Biol Chem. 1997;272:13816C13822. [PubMed]Goodnight JA, Mischak H, Kolch W, Mushinski JF. Immunocytochemical localization of eight protein kinase C isozymes overexpressed 1415562-82-1 in NIH 3T3 fibroblasts. Isoform-specific association with microfilaments, Golgi, endoplasmic reticulum, and nuclear and.