Oa hydrochloride

Female pheromone components were selected based on their ability to elicit single sensillum responses in female H. virescens moths [11 ]. The female sex pheromone components were as follows: (Z)-9-tetradecenal (Z9-14:Ald), (Z)-11-hexadecenal (Z11-16:Ald) and (Z)-11-hexadecenyl acetate (Z11-16:OAc) and were obtained from Bedoukian Research, Inc. (Danbury, Connecticut, USA). Putative host volatiles were selected from compounds that have shown physiological or behavioral effects on H. virescens in previous studies [11 ][12 ][39 (link)][40 (link)]: 2-phenyl ethanol, Z3-hexenol, along with racemic linalool and β-caryophyllene were obtained from Sigma Aldrich (St. Louis, MO, USA). All chemicals were diluted in hexane at decade steps from 10ng/μl-100μg/μl, and stored at -20°C. OA-hydrochloride was obtained from Sigma Aldrich (St. Louis, MO, USA) and stored at room temperature. OA was diluted to 50μg/μl in hemolymph Ringer solution [41 ] and stored in a fridge at 4°C.

Primers (Table S1) used in this manuscript were designed based on the predicted sequence of T. castaneum and synthesized by Invitrogen (Shanghai, China). The pGEM-T Easy Vector (Promega, Madison, WI, USA) was used to clone the PCR amplicon of TcOctβ2R. The expression vector pcDNA3.1(+) was a gift from Dr. Yoonseong Park of Kansas State University. High-quality plasmid DNA prepared by a QIAGEN Plasmid Plus Midi Kit (Hilden, Germany) was used for transient transfection and heterologous expression.
The HEK 293 cells were cultured adherently in a culture medium at 37 °C with 5% CO₂ content. The culture medium was composed of DMEM/F12 medium, 10% fetal bovine serum (FBS), 1% fungizone, and 1% penicillin/streptomycin. Coelenterazine h and the reagents used for cell culture were purchased from Gibco Life Technologies (Grand Island, NY, USA). The TransIT–LT1 transfection reagent used for the transient transfections was purchased from Mirus Bio Chemicals (Madison, WI, USA). OA hydrochloride, dopamine hydrochloride, TA hydrochloride, forskolin, and naphazoline hydrochloride were all purchased from Sigma-Aldrich (St. Louis, MO, USA). The GloSensor reagent used for the cAMP assay was purchased from Promega.