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. 2009 Mar 18;29(11):3551–3564. doi: 10.1523/JNEUROSCI.0415-09.2009

Figure 3.

Figure 3.

Thr-161 is required for cell surface expression of DOR and the formation of DOR–MOR heterodimers. A, No significant changes in the total expression level between wild-type DOR (WT) and T161A-DOR after transient transfection in NG108-15 cells. B, Surface-biotinylated DOR showed that the expression of functional DOR at the cell surface for the T161A mutant was significantly less than that of WT-DOR after transient transfection in NG108-15 cells. C, Cell lysates from NG108-15 cells cotransfected with EGFP-DOR (2 μg), HA-MOR (4 μg) cDNA, and Cdk5 siRNA or control siRNA were subjected to immunoprecipitation using polyclonal anti-GFP antibody. Associated HA-MOR was detected by immunoblotting using a monoclonal anti-HA antibody. The levels of DOR and MOR in each transfection are shown by immunoblotting with anti-GFP and anti-HA antibodies, respectively. D, Cell lysates from NG108-15 cells cotransfected with EGFP-DOR (wild type or T161A) (2 μg) and HA-MOR (4 μg) cDNA were subjected to immunoprecipitation using polyclonal anti-GFP antibody. Associated HA-MOR was detected by immunoblotting using a monoclonal anti-HA antibody. The levels of DOR and MOR in each transfection are shown by immunoblotting with anti-GFP and anti-HA antibodies, respectively. Results are representative of three independent experiments.