Tenascin c tnc

Following the generation of unpatterned or micropatterned methacrylate thin-films, adsorbed biochemical borders were created on the surface to investigate their role in neurite pathfinding on physical guidance cues. To create biochemical borders, equal volumes of solutions with different bioactive species, e.g. one solution containing laminin (20 µg/ml, Sigma-Aldrich) and the other containing tenascin-C (TnC, 100 µg/ml, Millipore) or EphA4-Fc (10 µg/ml, R&D Systems), were separately deposited on a methacrylate polymer surface overnight at 4 °C. A glass coverslip for each solution was placed on the dispensed volume so as to cover half of the polymer film while aligning adjacent to the other coverslip. Protein solutions subsequently spread across the surface due to capillary forces between the glass coverslip and the methacrylate thin-film forming a boundary between the two proteins where the two cover slips join. Following adsorption, polymer films were rinsed three times with phosphate-buffered saline (PBS).

UM-SCC 11b tumor line was obtained from the University of Michigan squamous cell carcinoma series in 2000 (Ann Arbor, MI). The FaDu (HTB-43™) and SCC-15 (CRL-1623) tumor lines were purchased from ATCC in 2000 (Manassas, VA) and were cultured as per instructions from ATCC or as previously described (23 (link)); newly received cells were expanded, frozen back. Cells were cultured for less than 3 months per thaw. Cell lines were not authenticated by authors. Tissue culture plasticware and nanofibers (Corning) were coated with 0.5 µg/cm² of 2:1 (w/w) Tenascin-C (TNC, Millipore):Fibronectin-1 (FN1, Sigma-Aldrich) in 1X phosphate buffered saline (PBS). DOTAP (N-[1-(2,3-Dioleoyloxy)propyl]-N,N,N-trimethylammonium methyl-sulfate) transfection reagent (Roche Applied Sciences) was used according to manufacturer’s protocol. PEI-polyplexes (25 kDa PEI, Sigma-Aldrich) were prepared and used as described (33 (link)). Nanocapsule transfections were performed as described in Figs. 1–4 legends.