FIG. 3.

Nm23-H1 coimmunoprecipitates with EBNA1 in EBV-transformed LCLs. LCL1 and LCL2 are recently transformed human B-cell lines by EBV. All of the latent genes are expressed, and they are both type III latency cell lines. Anti-Nm23-H1 antibody was used to incubate with lysates from 50 million cells for 12 to 16 h at 4°C. All complexes were collected by using protein A-Sepharose and fractionated on a SDS-12% PAGE gels. (A) Results when membranes were probed with antibodies against EBNA1 showing that EBNA1 coimmunoprecipitates in both cell lines with anti-Nm23-H1 antibodies. No signal was seen in the control EBV-negative cell line BJAB. (B) Results from a pull-down experiment with GST-Nm23-H1 fusion proteins conjugated to GST protein. Glutathione beads were bound to the fusion proteins and then incubated with lysates from the two LCLs. Complexes were again transferred to nitrocellulose membranes, and the membranes were probed with anti-EBNA1 antibodies. The results show signals for EBNA in the GST-Nm23H1 lanes (shown as Nm23-H1).