Investigating Taste Receptor Signaling in Smooth Muscle Cells

Dextromethorphan hydrobromide, chloroquine, denatonium benzoate, quinine hydrochloride, phenylthiocarbamide (PTC), 6-n-propylthiouracil (PROP), sodium thiocyanate, salicin, nicotine, yohimbine, colchicine, thiamine, caffeine, chloramphenicol and picrotoxinin were purchased from Sigma. Fluo-4 NW calcium assay kit was purchased from Invitrogen. hPASMCs and hASMCs were either purchased from ATCC or were a kind gift from Dr. Andrew Halayko, Dept. of Physiology, University of Manitoba. Cell culture media and hPASMC growth kit were purchased from Cedarlane, Canada. Nucleofection kit for smooth muscle cell transfection was purchased from Lonza. Polyclonal antibodies specific for T2R1, T2R38 and normal IgG specific rabbit antisera were obtained from Abcam (Cambridge, MA, USA) and Santa Cruz Biotechnology (Dallas, TX, USA) respectively [6] (link), [7] (link). Phospho-MLC-Ser19 monoclonal antibody and MLC polyclonal antibody were purchased from Cell Signaling technology (Danvers, MA, USA). DyNazyme hot start was purchased from Thermo Fisher Scientific (Toronto, ON, Canada). The shRNA specific for T2R1 was purchased from Qiagen (Toronto, ON, Canada).