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. 2020 Oct 6;11:574082. doi: 10.3389/fphys.2020.574082

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Copyright © 2020 Kurtz, Steinberg, Betcher, Fowler and Shepard.

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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Functional surface expression was achieved for 5 murine olfactory receptors. Hepatic olfactory receptors were cloned into a vector containing several N-terminal tags: Lucy and Rho to optimize surface expression and Flag for detection purposes. Receptors were transfected into HEK293T cells either alone or in combination with trafficking proteins RTP1s and/or Ric8b, and then live-surface labeled with a polyclonal Flag antibody to detect the protein expressed on the cell surface (Surface Flag). Cells were then fixed, permeabilized, and stained with a monoclonal Flag antibody to detect all transfected cells (Total Flag). DAPI was used to visualize nuclear staining. Scale bar = 20 μm.