Figure 6.
WT and R295S NK3Rs dimerize and both receptors interact with Gαq protein. A) COS-7 cells coexpressing V5-tagged WT NK3R and HA-tagged WT or R295S (RS) NK3R were lysed and lysates were used in a coimmunoprecipitation (IP) assay. Proteins were immunoprecipitated with an anti-V5 antibody (right panel) and subjected to Western blot analysis and immunoblotting (IB) with an anti-HA (top panels), anti-V5 (middle panels), or anti-β-actin (bottom panels) antibody. Fifteen percent of the lysates were used as input and subjected directly to immunoblotting (left panels). B) Cell lysates from COS-7 cells expressing WT or R295S NK3R, or both, were prepared, and immunoprecipitation was performed with an anti-Gαq/11 antibody coupled to agarose G beads (right lanes). Immunoprecipitated proteins were subjected to Western blot analysis and immunoblotted with an anti-HA antibody (top panel) or an anti-β-actin antibody (bottom panel). One percent of the lysates was used as input and subjected directly to immunoblotting (left lanes).