Figure 4.

RORα recruits NcoR1 instead of p300 to ERRγ in the CYP2E1 promoter. (A) The primary cultures of mouse hepatocytes were treated with 20 μM ACEA or 20 μM CS for 24 h as indicated. (B) The primary cultures of mouse hepatocytes were transfected with si-GFP or si-RORα, and treated with 20 μM ACEA and/or 20 μM CS as indicated. Whole cell lysates were IP and probed using the indicated antibodies by WB. The level of proteins in the cell lysates was analyzed by WB as input. (C) Chang liver cells were transfected with the ERRE-Luc with empty vector, the expression vectors encoding FLAG-ERRγ, FLAG-RORα or increasing amount of MYC-p300 for 24 h as indicated (left). Or cells were transfected with the ERRE-Luc with the receptor expression vectors in the presence of si-NCoR1 for 48 h as indicated (right). Expression of NCoR1 protein after si-RNA-mediated knock-down was shown in Supplementary Figure S8. (D) Chang liver cells were transfected with the ERRE-Luc with the expression vectors encoding ERRγ, RORα or NCoR1 for 24 h as indicated. Luciferase activity that normalized to the corresponding β-galactosidase activity was converted to fold activity with no treatment as 1. The data represent mean ± SD. *P < 0.05 versus EV; ^#P < 0.05 versus ERRγ; ^≠P < 0.05 versus ERRγ with RORα (n = 3). (E) Whole cell lysates obtained from the Chang liver cells transfected with FLAG-ERRγ, ERRγ-ΔAF2, MYC-RORα and/or RORα-ΔAF2 were IP and probed using the indicated antibodies by WB. The level of proteins in the cell lysates was analyzed by WB as input. Representatives of at least three independent experiments with similar results are shown.