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. 2019 Feb 18;10:92. doi: 10.3389/fphys.2019.00092

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Copyright © 2019 Pazo, Pérez-González, Oliveros, Huarte, Chavez and Nieto.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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hCLE forms oligomers. (A) Total protein extracts of HEK293T cells transfected with plasmids expressing the whole hCLE sequence (hCLEwt), N-terminal domain [hCLE(Nt)] (aa 1–120) or C-terminal domain [hCLE(Ct)] (aa 120–244) were analyzed by Western blot with anti-hCLE antibodies. (B) Silver staining of hCLE-CBD purified protein dyalized in the presence of non-reducing agents (No. Reduc.). (C) Purified hCLE was dyalized in reducing (Reduc.) or non-reducing buffer (No Reduc.) and analyzed in non-denaturing gel followed by Western blot with anti-hCLE antibodies. ^∗monomer and ^∗∗dimers. 1X and 2X represent single or double amount of purified protein. Three independent experiments were done.