Extended Data Fig. 4: Influence of gradient transfection of GEM on cAMP accumulation and β-arrestin 2 recruitment mediated by D1R, and the effects of GEM insertions on dopamine-induced D1R signaling.

a-h, Dopamine dose-response curves for cAMP accumulation with increasing amounts of \(\text{GE}{\text{M}}_{\text{anchor}}^{\text{TM}1/2/4}\) (a), \(\text{GE}{\text{M}}_{\text{BAM}}^{\text{TM}3/4/5}\) (b), \(\text{GE}{\text{M}}_{\text{NAM}}^{\text{TM}5/6/7}\) (c) and \(\text{GE}{\text{M}}_{\text{ago}-\text{PAM}}^{\text{TM}5/6/7}\) (d) and for β-arrestin 2 recruitment with increasing amounts of \(\text{GE}{\text{M}}_{\text{anchor}}^{\text{TM}1/2/4}\) (e), \(\text{GE}{\text{M}}_{\text{BAM}}^{\text{TM}3/4/5}\) (f), \(\text{GE}{\text{M}}_{\text{NAM}}^{\text{TM}5/6/7}\) (g) and \(\text{GE}{\text{M}}_{\text{ago}-\text{PAM}}^{\text{TM}5/6/7}\) (h). i-l, Dose-response curves showing the effects of various D1R constructs in which \(\text{GE}{\text{M}}_{\text{anchor}}^{\text{TM}1/2/4}\) (i), \(\text{GE}{\text{M}}_{\text{BAM}}^{\text{TM}3/4/5}\) (j), \(\text{GE}{\text{M}}_{\text{NAM}}^{\text{TM}5/6/7}\) (k) and \(\text{GE}{\text{M}}_{\text{ago}-\text{PAM}}^{\text{TM}5/6/7}\) (l) were fused with the receptor. The signaling was dopamine-induced, measured by cAMP accumulation and β-arrestin 2 recruitment assays. Values are shown as the mean ± SEM from three independent experiments performed in triplicate. A detailed statistical evaluation is provided in Extended Data Tables 1–3.