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. 2010 Dec 30;5(12):e16000. doi: 10.1371/journal.pone.0016000

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Bohr et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A–D) Purified fraction of CCN3/NOV (A, B) or CCN2/CTGF (C, D) that were stored at indicated temperatures for three months were subjected to denaturing (+ DTT) and non-denaturing (− DTT) SDS-PAGE (Bis/Tris 4–12% gradient). The gels were blotted onto Protran membranes and subsequently probed with a CCN3/NOV- (A), a CCN2/CTGF- (C), or a ConA-HRP conjugate (B, D). In this analysis, a soluble PDGF type β receptor (PDGFRβ) was used as a highly glycosylated protein control. BSA and the bacterial expressed CCN3/CTGF (BioVendor) were used as negative controls. In this set of experiments the blots A and C were cut at indicated cut edges prior incubation with antibodies directed against CCN proteins or PDGFRβ.