Figure 3

Hedgehog signaling activity as reflected by Gli-Lux reporter transfection. Normalized luciferase activity of CD133⁺/EpCAM⁺ and CD133⁻/EpCAM⁻ subpopulations was evaluated by co-transfecting with the Gli-Lux in which firefly luciferase is under the control of the Gli promoter and pRL-TK luciferase reporter system in which renilla luciferase is under the control of the TK promoter. (a) Relative luciferase activity in CD133⁻/EpCAM⁻ subpopulation was determined after transfection with Gli-Lux and a mutated plasmid (Gli-Lux-M). **P<0.01 compared with CD133⁺/EpCAM⁺ subpopulation. Treatment with Hh signaling SMO antagonists, itraconazole (b), or LDE225 (c) resulted in a decrease in firefly luciferase activity in CD133⁻/EpCAM⁻ subpopulation. **P<0.01 compared with CD133⁻/EpCAM⁻ subpopulation without treatment.