BioMed Research International / 2010 / Article / Fig 2

Research Article

The Collagen V Homotrimer Production Is Unexpectedly Favored over the Heterotrimer in Recombinant Expression Systems

Figure 2

Biochemical analysis of the recombinant pro 𝛼 2(V) chains produced in HEK-293 cells. (a) 6% SDS-PAGE analysis of cell media from untransfected cells (lane 1) and from cells transfected with human pro 𝛼 2(V) construct (lanes 2–5) under unreduced (lanes 1, 2) or reduced (lanes 3, 4, 5) conditions. Transfected cells media were incubated without (lane 4) or with (lane 5) pepsin before SDS-PAGE analysis. (b) Digestion with bacterial collagenase: transfected cell media were incubated without (lane 1) or with (lanes 2, 3) collagenase before 12% SDS-PAGE analysis under reduced (lanes 1, 2) or unreduced (lane 3) conditions. (c) Digestion with BMP-1: 12% SDS-PAGE patterns of transfected cells media digested (lane 2) or not (lane 1) with BMP-1. Gels (a) and (b) were stained with Coomassie blue whereas gel (c) was silver stained. Left, molecular mass standards expressed in kDa. NT: not transfected, pro 𝛼 2(V): transfected with pro 𝛼 2(V) construct, DTT: dithiothreitol, Cpro: C-propeptide, Npro: N-propeptide, BMP-1: Bone Morphogenetic Protein 1.

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