Figure 4.

AT1R couples to Gαi within the AT1R–CB₁R heteromer. (A) Neuro2A-AT1R cells transfected or not with a siRNA to CB₁R (siCB₁R) in 24-well plates were incubated with increasing concentrations of Ang II in the absence or presence of the CB₁R antagonist PF514273 (1 μM) or (B) Neuro2A and Neuro2A-AT1R cells in 24-well plates were incubated with increasing concentrations of Hu210 at 37°C for 15 min in the cAMP treatment buffer (0.5 mM isobutylmethylxanthine and 10 μM forskolin in Krebs-Ringer-HEPES buffer). After terminating the reaction by heating at 95°C, cAMP concentrations were determined as described in ‘Materials and methods’. Data are expressed as the mean±s.e.m. (n=3 independent experiments). ^***P<0.001. (C) Data are plotted to compare the inhibition of cAMP production across the different experimental conditions. Data are expressed as the mean±s.e.m. (n=3 independent experiments). ^***P<0.001, indicated conditions versus Ang II; ###P<0.001, Hu210 treatment of Neuro2A versus Hu210 treatment of Neuro2A-AT1R.