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Membrane expression of receptors

JL Jenny Landin
DH Daniel Hovey
BX Bo Xu
DL David Lagman
AZ Anna Zettergren
DL Dan Larhammar
PK Petronella Kettunen
LW Lars Westberg
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HEK293 cells were grown on coverslips treated with poly-D-lysine. The expression constructs of receptors, with GFP tagged on the C-terminal, were transfected using Lipofectamine 2000 (Thermo Fisher Scientific), then incubated for 24 hours at 37 °C, with 5% CO2. The medium was discarded and the coverslip with HEK293 cells was gently washed twice with PBS. Cells were then fixed with 95% ethanol for 10 minutes, and following this, coverslips were placed upside down onto slides containing a drop of Vectashield mounting medium (with DAPI), and sealed with nail polish. GFP expression pictures were produced using a confocal microscope.

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