Fig. 6.

TRAF2 is required for Na-mediated induction of lytic protein expression. (A) Hone-Akata cells were transfected with control or TRAF2 siRNAs. At 48 h after transfection, the cells were retransfected with the control or TRAF2 siRNAs along with control or FLAG-Na expression plasmids. Immunoblot analysis was performed 2 days later using antibodies against BMRF1 and Z to examine lytic protein expression, or with TRAF2 and FLAG to detect TRAF2 and Na, respectively. β-Actin was used as a loading control. (B) CNE2-Akata cells were transfected with lentiviruses expressing shRNAs against either a control sequence or TRAF2. At 48 h posttransfection, cells were retransfected with the shRNA vectors along with either control or FLAG-Na expression plasmids. Cell lysates were harvested, and immunoblot analyses were performed using antibodies against TRAF2, BMRF1, FLAG (Na), R, Z, and β-actin. The data shown represent samples run on the same gel; irrelevant lanes were cropped from the figure. (C) Stable selected Hone-Akata cell lines transduced with lentiviruses expressing shRNAs against a nonspecific control or TRAF2 were transfected with either vector control or FLAG-Na expression vectors as described above. Immunoblot analysis was performed using antibodies against phosphorylated JNK (p-JNK), total JNK, BMRF1, TRAF2, FLAG (Na), and β-actin.