*4.5. Cell Culture*

Shh-Light II cells (JHU-068) were maintained in Dulbecco's Modified Eagle Medium (DMEM) (Gibco, Carlsbad, CA, USA) supplemented with 0.4 mg/mL geneticin, 0.15 mg/mL Zeocin™ (Invitrogen, Carlsbad, CA, USA), and 10% bovine calf serum. The cells were grown at 37 ◦C in an atmosphere of 5% CO2 in air and 100% relative humidity. This mouse embryonal NIH 3T3 cell line contains a stably transfected luciferase reporter with eight copies of the consensus Gli binding site [35]. Alkaloid treatment conditions were dissolved in ethanol and added to DMEM media containing 0.5% bovine calf serum.
