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. Author manuscript; available in PMC: 2016 Dec 1.
Published in final edited form as: Cell Metab. 2015 Sep 17;22(6):1009–1019. doi: 10.1016/j.cmet.2015.09.003

Figure 3. REV-ERBa and β are necessary for MYC disruption of BMAL1 promoter oscillation.

Figure 3

A. mRNA levels (normalized to expression of β2M) for PER2 or CRY1 determined by RT-PCR in U2OS MYC-ER cells transfected with either non-targeting siRNA (Con), siRNA against PER2, or CRY1 for 48 h with 4OHT (MYC-ON) being added to cells 24 hours after transfection. Means and SDs from at least three experiments are shown. *p < 0.05 for Con vs. siPER2 or siCRY1. mRNA (FC) = Fold Change. B. BMAL1 mRNA expression (normalized to expression of β2M) in U2OS MYC-ER transfected as described in (A) with siRNAs indicated at the bottom. *p < 0.05 by Student’s t test for MYC-ON vs. MYC-OFF. ns = not significant. C. U2OS MYC-ER cells were transfected with siRNAs as indicated in the figure, synchronized with dexamethasone and monitored using a LumiCycle™ luminometer. Data are representative of two or more experiments. D. mRNA levels (normalized to expression of β2M) for REV-ERBα or REV-ERBβ determined by RT-PCR in U2OS MYC-ER cells transfected with either non-targeting siRNA (Con), siRNA against REV-ERBα, REV-ERBβ, or both REV-ERBs (α+β) for 48 h with 4OHT (MYC-ON) being added to cells 24 hours after transfection. Means and SDs from at least three experiments are shown. *p < 0.05 for siREV vs. control. E. BMAL1 mRNA levels (normalized to expression of β2M) in U2OS MYC-ER transfected with the indicated siRNAs (bottom) as described in (D) and cultured with 4OHT (MYC-ON) or EtOH (MYC-OFF) after 24 hours. Means and SDs from at least three experiments are shown. *p < 0.05 by Student’s t test for siREV or Con MYC-ON vs. Con MYC-OFF. §, p< 0.05 siREV vs. Con MYC-ON. F. U2OS MYC-ER cells transfected with the indicated siRNAs for 48 hours, cultured with 4OHT (MYC-ON) or EtOH (MYC-OFF), synchronized with dexamethasone and monitored as in (C). Data are representative of two or more experiments. RLU = relative light units.