FIG 2.

EBNA3C attenuates H2AX levels in Burkitt's lymphoma and epithelial cells. (A) Approximately 30 million BJAB, DG75, and Ramos cells were harvested for WB analysis of H2AX, EBNA3C, GAPDH, and GFP levels after transient transfection of EBNA3C and GFP constructs. (B) Endogenous expression of H2AX levels in Burkitt's lymphoma cell lines transfected with EBNA3C was analyzed for transcript levels by real-time PCR. Approximately 15 million BJAB, DG75, and Ramos cells were cotransfected with the GFP-expressing vector with either EBNA3C or the control vector. Transcript levels of the H2AX gene are represented as fold changes. The experiments were performed in triplicate. (C) H2AX in HEK293 cells transfected with EBNA3C in a dose-dependent manner was analyzed at the transcript level by RT-PCR. Transcript levels of the H2AX gene are represented as fold changes from experiments performed in triplicate. (D) H2AX in HEK293 cells transfected with EBNA3C in a dose-dependent manner was analyzed at the protein level. (E) H2AX and EBNA3C constructs were transfected into HEK293 cells and analyzed for the detection of H2AX protein levels. HEK293 cells were transfected with GFP, EBNA3C-Flag, or the vector with Myc-H2AX. Data in all panels are representative of repeat experiments.