The Collagen V Homotrimer Production Is Unexpectedly Favored over the Heterotrimer in Recombinant Expression Systems
Figure 3
Selection of clones expressing 1(V) and 2(V) chains. Western blot analysis with antibody to collagen V of recombinant 1(V) and 2(V) chains synthesized and secreted in the culture medium of co-transfected HEK-293 cells. The constructs 1(V)-pZeo-SV2 (lanes 2, 3) or 1(V)-pcDNA-Hygro (lane 4) and the construct 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 1(V)/2(V): clone C22, which expressed more of the 2(V) chain than the 1(V) chain, clone C32, which expressed both chains at approximately the same amount, and clone F22, which expressed more of the 1(V) chain than the 2(V) chain. Left, molecular mass standards expressed in kDa.
