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. 2015 Jul 23;6(28):25784–25800. doi: 10.18632/oncotarget.4703

Figure 2. Effect of AF4, MLL, AF4-MLL and MLL-AF4 overexpression on VDR/RXR- and calcitriol/TGFβ-dependent response of the reporter gene constructs A. pN10 B. pN10cdsInJM and C. pGL3cdsInJM.

Figure 2

In order to study inducibility of the constructs by calcitriol/TGFβ, HeLa cells were co-transfected either with the pSG5 control vector or with the expression vectors for the nuclear receptors VDR and RXR. 16 hours after transfection, cells were incubated without or with calcitriol (50 nM) and TGFβ (1 ng/ml). After 40 hours, luciferase activity was determined. Results are given as RLU (left panels) which include normalization to transfection efficiency and as x-fold induction by VDR/RXR over pSG5 control (middle panels) or x-fold induction by calcitriol/TGFβ over VDR/RXR (right panels). Each experiment was performed in triplicate. Results are presented as mean + S.E.M. of three independent experiments. Two-tailed t-test was performed to determine significant differences in relation to co-transfection of the backbone vector pTarget, indicated by asterisks (*P < 0.05, **P < 0.01, ***P < 0.001).