Fig. 1
Expression properties and molecular interaction of neurokinin receptors (NKs). A RT-PCR analysis of mRNA expression. Total RNAs from cell lines were analyzed with isotype-specific primers and PCR products after 35 cycles for the receptors or 25 cycles for β-actin gene were visualized on a 1.5% agarose gel. A 1 kb-plus ladder was used as a size marker. B N-terminal HiBiT-tagged receptors were expressed in HEK293 cells with varying amounts of plasmids. HiBiT activity was assessed by adding Nano-Glo® HiBiT extracellular detection reagent. **: p < 0.01 vs. vector C Expression of C-terminal HA-tagged NKs. HEK293 cells transfected with plasmids containing C-terminal HA-tagged NKs were lysed with RIPA buffer and subjected to western blotting with anti-HA antibodies. D Luminescence induced by receptor dimerization. HEK293 cells co-expressing NKs of C-terminal SmBiT- and LgBiT-tagged forms were analyzed by NanoBiT assay. **: p < 0.01 vs. vector (V). E Co-immunoprecipitation. HEK293 cells co-expressing FLAG- and HA-tagged receptors were lysed and subjected to immunoprecipitation with anti-FLAG agarose, followed by western blotting with anti-HA antibody