20α-Hydroxy Cholesterol is an allosteric activator of the Hedgehog signaling pathway Smoothened (Smo) oncoprotein that binds at a site distinct from the canonical cyclopamine binding site. 20α-Hydroxy Cholesterol activates Hedgehog (Hh) signaling (EC50 ~ 3μM for induction of Hh reporter gene transcription in in NIH 3T3 cells). It induces Smo accumulation in primary cilia and also exhibits osteogenic activity and activates the liver X receptor (LXR).