COS-7 cells stably expressing hY1/2/4/5R_eYFP fusion protein and the Δ6Gαqi4myr chimeric Gα-protein were prepared as previously described [27 (link)]. The hY1/2/4/5R_eYFP cDNA was subcloned into MCS1 of a pVitro2-MCS vector carrying a hygromycin resistance gene; the Δ6Gαqi4myr cDNA was subcloned into MCS1 of a pVitro2-MCS vector carrying a G418 resistance gene (Invivogen, San Diego, CA).
COS-7 (African Green Monkey kidney) cells stably expressing the hY1/2/4/5R_eYFP fusion protein and the Δ6Gαqi4myr chimeric Gα-protein were cultured at 37°C in high glucose Dulbecco’s Modified Eagle Medium (DMEM, Life Technologies, Carlsbad, CA) with glutamine and sodium pyruvate (Life Technologies/Lonza) supplemented with 10% (w/v) FBS (Invitrogen, Carlsbad, CA), 1.5 mg/mL G418-sulfate (Amresco, Solon, OH) and 133 μg/ml hygromycin (Invivogen).
COS-7 (African Green Monkey kidney) cells stably expressing the hY1/2/4/5R_eYFP fusion protein and the Δ6Gαqi4myr chimeric Gα-protein were cultured at 37°C in high glucose Dulbecco’s Modified Eagle Medium (DMEM, Life Technologies, Carlsbad, CA) with glutamine and sodium pyruvate (Life Technologies/Lonza) supplemented with 10% (w/v) FBS (Invitrogen, Carlsbad, CA), 1.5 mg/mL G418-sulfate (Amresco, Solon, OH) and 133 μg/ml hygromycin (Invivogen).
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