The EBV Latent Antigen 3C Inhibits Apoptosis through Targeted Regulation of Interferon Regulatory Factors 4 and 8
Figure 7
EBNA3C differentially modulates poly-ubiquitination of IRF4 and IRF8.
A, B, C, D) 10 million HEK-293 cells were transiently transfected with different combinations of expression vectors as indicated. Cells were harvested after 36 hrs of post-transfection and total protein was immunoprecipitated (IP) with indicated antibodies and protein samples were resolved by 10% SDS-PAGE. Western blots were performed by stripping and re-probing the same membrane. E) 50 million EBV negative BJAB cells, BJAB cells stably expressing EBNA3C (BJAB7, BJAB10) and EBV transformed LCL1, LCL2 were incubated with proteasome inhibitor MG132 drug (20 µM) for 6 hrs. Cells were harvested and lysed with RIPA buffer. IRF4 was immunoprecipitated (IP) by using specific antibodies. Samples were resolved by 10% SDS-PAGE. Western blotting (WB) was performed by stripping and reprobing the same membrane. F–G) Ubiquitination assay was performed with Lentivirus mediated stable EBNA3C knockdown (Sh-E3C) or scramble control (Sh-Ctrl) LCL1 cells and subjected to Western blot analysis with indicated antibodies.