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. 2019 Dec 20;14(12):e0226570. doi: 10.1371/journal.pone.0226570

Fig 1. Evaluation of Wnt, Hh and Notch luciferase single reporters via luminescence readout.

Fig 1

(A) The Wnt promoter TOP was combined with NLuc (substrate: furimazine). (D) The Hh promoter GLI-RET was backbone switched to obtain GLI-FLuc (substrate: luciferin). (G) For the Notch (Not) reporter construct, the CBF promoter was combined with GLuc (substrate: coelenterazine). All transfections were carried out in 293T cells. B, E, H: Luminescence measurement of cells transfected with the indicated plasmid and co-transfected with either pUC19 control or the indicated inducer plasmid. 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 show from a representative experiment; C, F and I are the average of 2–5 independent experiments. (**P≤0.01, ****P≤0.0001, ns = not significant, t-test, n = 4). Normalised RLUs were multiplied with 10 (CBF-GLuc), 100 (TOP-NLuc) or 1000 (FOPFlash, TOPFlash, GLI-RET and GLI-FLuc).