Figure 3.

Switch in AT1R G protein coupling to the ERK pathway within the AT1R–CB₁R heteromer. Neuro2A-AT1R cells were starved for 4 h and the levels of pERK were measured after various treatments (see below), after 3 min stimulation with 10 nM Ang II. (A) Phospho-ERK levels after Ang II stimulation were examined in Neuro2A-AT1R cells, after transfection with a Gαq dominant-negative construct (DN Gαq, see insert). (B) Phospho-ERK levels after Ang II stimulation were examined in Neuro2A-AT1R cells transfected with a siRNA to CB₁R, after transfection with or without a Gαq dominant-negative construct. (C) Data are expressed as the mean±s.e.m. (n=4 independent experiments). ^*P<0.05; ^***P<0.001. (D) Phospho-ERK levels after Ang II stimulation were examined in Neuro2A-AT1R cells after incubation with pertussis toxin (PTX; 15 ng/ml for 16 h). (E) Phospho-ERK levels after Ang II stimulation were examined in Neuro2A-AT1R cells transfected with a siRNA to CB₁R, after incubation with PTX. (F) Data are expressed as the mean±s.e.m. (n=4 independent experiments). ^*P<0.05; ^***P<0.001.