EBNA3C specifically down-regulated RASSF1A protein expression.

(A-B) RASSF1A expression was decreased in EBV positive cell lines. 15 million A) Burkitt’s lymphoma cell BL41 and Akata B) Mutu I and Sav I cells and corresponding EBV positive A) BL41-B95.8 and Akata-EBV B) Mutu III and Sav III cells were harvested and lysed with RIPA buffer. The expression levels of RASSF1A were detected by western blot. The relative density (RD) of RASSF1A was quantitated and shown. (C-D) RASSF1A expression was repressed in EBNA3C stably expressing BJAB cells, and EBV-transformed LCLs. The expression of RASSF1A in C) BJAB7 and BJAB10 and D) LCL1 and LCL2 cells were detected as described for A. (E-G) EBNA3C decreased RASSF1A expression specifically. E) Saos-2, F) 293 and G) BJAB cells were transfected with Myc-tagged RASSF1A alone or co-transfected with an increasing amount of Flag-tagged EBNA3C. Western blots were performed to detect the expression of RASSF1A as described for A. (H-I) Knocking down EBNA3C rescued the expression level of RASSF1A in BJAB10 and LCL cells. H) 10 million BJAB10 cells were transfected with sh-EBNA3C or scramble control (sh-Ctrl) by electroporation. I) 15 million lentivirus transduced stable EBNA3C knocked-down LCL1 cells and LCL1-sh-Ctrl were harvested. 48h post-transfection, the cells were harvested and lysed for western blot. The relative density (RD) of RASSF1A was quantitated and shown.