BioMed Research International / 2010 / Article / Fig 3

Research Article

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

Figure 3

Selection of clones expressing ๐›ผ 1(V) and ๐›ผ 2(V) chains. Western blot analysis with antibody to collagen V of recombinant ๐›ผ 1(V) and ๐›ผ 2(V) chains synthesized and secreted in the culture medium of co-transfected HEK-293 cells. The constructs ๐›ผ 1(V)-pZeo-SV2 (lanes 2, 3) or ๐›ผ 1(V)-pcDNA-Hygro (lane 4) and the construct ๐›ผ 2(V)-Rc-CMV (lanes 2, 3, 4) were co-transfected into the HEK-293 cells. These cells do not naturally express these chains (lane 1). Three clones have been selected to express different ratios of ๐›ผ 1(V)/ ๐›ผ 2(V): clone C22, which expressed more of the ๐›ผ 2(V) chain than the ๐›ผ 1(V) chain, clone C32, which expressed both chains at approximately the same amount, and clone F22, which expressed more of the ๐›ผ 1(V) chain than the ๐›ผ 2(V) chain. Left, molecular mass standards expressed in kDa.

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