Fig. 3. The three macrostates divided by MSM, and constitutive activity.

A The distribution of active (green), inactive (yellow), and intermediate (purple) states on the free-energy landscape. The attribution and probability of each macrostate are shown on the right. B The transition time among the active (green), inactive (yellow), and intermediate (purple) states, represented by the mean first passage time. Constitutive activities of AT₁ receptor in Gq (C), Gi (D), and G12 (E) pathways (red). Vasopressin 2 receptor (V₂R) was used as a negative control (black). The gradient cell surface expression levels of AT₁ receptor and V₂R were achieved by adjusting the transfecting amounts of plasmids encoding the respective receptor in HEK293 cells (Supplementary Fig. 3). Data were from three independent experiments. The bars indicate the mean ± SEM values. The absolute luminescence intensity values are provided in source data. If no special instructions were shown, histograms were drawn by GraphPad. ΔBRET: the change of bioluminescence resonance energy transfer value.