Fig. 4.

Overexpression of Nrf2 up regulates endogenous and transfected Bcl-xL gene expression. (A). HEK-293 and HEK-293-Nrf2 cells expressing tetracycline-induced FLAG-tagged Nrf2 were treated with 2 µg/ml tetracycline for the indicated times. Sixty µg cell lysates were immunoblotted with anti-FLAG, anti-Bcl-xL, anti-NQO1 and anti-actin antibodies (upper panel). The band intensities of Bcl-xL and NQO1 from “A” were quantified and presented below the blots. (B & C). HEK293 and HEK293-Nrf2 cells were co-transfected with wild type pGL2B-Bcl-xL-1.565-WT-LUC or mutant type pGL2B-Bcl-xL-1.565-MT-LUC with the internal control Renilla luciferaseplasmid pRL-TK (B) or co-transfected with pGL2p-ARE-F1-LUC or pGL2p-mutant ARE-F1-LUC plasmids with the internal control Renilla luciferase plasmid pRL-TK (C). pGL2B and pGL2p vectors were also transfected as negative control. Twenty-four hours after transfection the cells were treated with 2µg/ml tetracycline for 16h and lysates were analyzed for luciferase activity. The data shown are mean ± S.D. of three independent transfection experiments.