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Collagen processing determines the sheet forming capacity in AF clones.

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posted on 28.01.2016, 12:39 by Guus G. H. van den Akker, Don A. M. Surtel, Andy Cremers, Stephen M. Richardson, Judith A. Hoyland, Lodewijk W. van Rhijn, Jan Willem Voncken, Tim J. M. Welting

A) Immunoblot analyses of Collagen type I protein in AF-S and AF-nS clones cultured in control or TGFβ3 containing medium for 7 days. Procollagen-alpha 1 (180 kDa; pro-α1) and Procollagen alpha 2 (145 kDa; pro-α2) variants of Collagen type I are indicated. The appearance of mature alpha 1 (135 kDa; α1) and alpha 2 (120 kDa; α2) variants of Collagen type I correlated well with sheet formation in AF-S clones. β-Actin (βACT) was used as loading control. B) Immunoblot analyses of Collagen type I protein in AF-S clones cultured in TGFβ3 medium with or without ascorbic acid for 7 days. Alpha-tubulin (αTUB) was used as loading control. C) Quantification of Collagen maturation as a function of time in AF-S and AF-nS clones. The ratio of mature COL1A1 over ProCOL1A1 and mature COL1A2 over ProCOL1A2 are depicted in the left and right graphs, respectively. At t = 0 no mature forms of Collagen type I were detectable. Statistical significance was assessed by Student’s t-test; * p<0.05.

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