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. 2007 Nov 8;149(2):695–702. doi: 10.1210/en.2007-0779

Figure 1.

Figure 1

EGF stimulates Dio2 mRNA and activity in JEG3 cells. A, Time course of EGF-treated JEG3 cells. Cells were incubated overnight with serum-free medium and then treated with 100 ng/ml EGF for the indicated times. RT-PCR was performed using primers specific for Dio2 and β-actin. B, Quantitative real-time PCR. Cells were treated as described above for 2 h with EGF. SYBR-Green quantitative PCR was performed using primers specific for human Dio2 and GAPDH. *, P < 0.05, EGF vs. control, Mann-Whitney U test. C, Effect of EGF treatment on D2 enzymatic activity. D2 assays were performed on sonicates of JEG3 cells stimulated for 6 h with EGF. *, P < 0.05, EGF vs. control, Mann-Whitney U test. D, Luciferase assay of 1.3-kb Dio2 Luc promoter construct. Cells were seeded in six multiwell plates and transfected with the 1.3-kb Dio2 Luc and RSV β-gal reporter plasmids. The day after transfection, cells were incubated for the indicated times with EGF. At the end of the incubation, luciferase and β-galactosidase assays were performed. *, P < 0.05, EGF vs. control, Mann-Whitney U test. Results (B–D) represent the average value ± sd of at least three different experiments, each performed in triplicate.