Panels (A–B) (top): Cholesterol occupancy time per residue of β2AR described in terms of the normalized time fraction, where a value of one stands for a contact throughout the simulation trajectory and zero means no contact. Results are given for (A) 10 and (B) 40 mol% of cholesterol. The residues of β2AR are defined to be in contact with cholesterol when any non-hydrogen atom of the residue is within ≤0.5 nm of any heavy atom of cholesterol. The data show that there are several hot spots (blue) as cholesterol binding sites. These plots were averaged from all independent simulations for a given cholesterol concentration, where the equilibration time (the first 100 ns of the simulation) was disregarded from the analysis. Panels (C) (bottom): Interaction sites as obtained from our simulations, are shown from two perspectives around the protein. EC and IC stand for extracellular and intracellular, respectively. Interaction sites at the intracellular (IC) side: IC1 (dark green) between helices (H) 1–4, IC2 (red) between H5 and H6, IC3 (magenta) between H3 and H5, and IC4 (orange) between H1 and H8. Interaction sites at the extracellular (EC) side: EC1 comprised of two closely placed cholesterol molecules between H5 and H6 (cyan) and in space surrounded by H6-ECL3-H7 (green), EC2 (purple) between H3 and H4, and EC3 (blue) between H1-H2-ECL1 (where ECL stands for the extracellular loop).