(A) The Wnt promoter TOP was combined with iRFP. (D) The Hh sensitive promoter PTCH1 (wildtype promoter for the hedgehog receptor PTCH1 [30]) was joined with mTurquoise2 (mT2). (G) For the Notch (Not) reporter construct, the CBF promoter was combined with tdTomato (tdT). Transfection were carried out either in 293T cells (B, C, E, F) or HeLa (H, I). B, E, H: Fluorescence measurement of cells transfected with the indicated plasmid and co-transfected with either pUC19 control (white bars) or the indicated inducer plasmid (grey bars). C, F, I: fold increase in signal for the indicated plasmid upon co-transfection with pUC19 control versus inducer plasmid (C: pWnt3a; F: phGli1; I: phICN1). In B, E and H results are shown from a representative experiment; C, F and I are the average of 3–6 independent experiments. (**P≤0.01, ****P≤0.0001, ns = not significant, t-test, n≥4).