Figure 2.

LMP1 and LMP2A activate Nrf2 in EBV-transformed B cells. EBV-transformed B cells were transfected with siRNA against LMP1 or LMP2A for 48 hours. (A) Whole cell extract and (B) cytosolic and nuclear extracts were prepared from siRNA-LMP1 or LMP2A-transfected cells. The protein levels of Nrf2 were measured by Western blot analysis. β-Actin and Lamin B1 were used as equal loading controls for normalization. CE, cytosol extract; NE, nuclear extract. The fold increase in LMP1, LMP2A, and Nrf2 is indicated numerically, as determined by densitometry. (C) The expression level of Nrf2 was detected by immunofluorescence. DAPI was used to counter stain the nucleus. Photographs were taken at 400× magnification. (D) ROS were detected with carboxy-H₂DCFDA. The data are expressed as the mean ± S.D. *P < .001. (E) The protein levels of HO-1 and NQO-1 were measured by Western blot analysis. The fold increase in NQO-1 and HO-1 is indicated numerically, as determined by densitometry. The experiments were performed in triplicate.