The SMO receptor is a class F GPCR and a vital component of the Hedgehog signaling cascade responsible for embryonic development and the regulation of adult stem cells . Like β2AR and other GPCRs, it is suggested that SMO is regulated via interactions of its transmembrane domain with cholesterol. A study by Hedger et al. has recently identified a cholesterol interaction site using a multiscale MD approach that combined Martini 2 and GROMOS53a6 84 . This interaction site is located on the transmembrane domain of SMO at the extracellular tips of helices H2 and H3 (Figure a). Our model recognized this interaction site (BS1) on the surface of SMO (Figure a, b, and c) in agreement with the findings of Hedger et al. . A second interaction site (BS2) was also observed between H1 and H7 (Figure a, b, and d) with similar residence time as the one described by Hedger et al. VDAC1 is a voltage-dependent anion channel with a number of binding sites for cholesterol. It is responsible for the regulation of ion and metabolite flux between the cytosol and the mitochondrial intermembrane space . Several theoretical and experimental studies have identified numerous cholesterol interaction sites on the transmembrane surface of VDAC1. Our Martini 3 cholesterol model binds to VDAC1 via 3 distinct binding sites (Supplementary Figure ). Binding sites 1 and 2 match well with some of the cholesterol-binding sites detected with docking procedures (BS1 with DS1, BS2 with DS4, and BS3 with DS5) 135 . The three binding sites are also in rough agreement with those detected on the surface of VDAC1 using Martini 2 . It is worth noting that we did not detect significant cholesterol occupancy on some of the binding sites predicted by docking, namely DS2 and DS3 135 . These were also not significantly populated by the previous Martini 2 model . Finally, binding site 3, the binding site with higher residence time detected, is well in agreement with a cholesterol-binding site detected by photo-affinity labeling experiments 136,137 .