Figure 2

Protein levels and DNA binding. a) Western blot showing EBNA-1 and Oct-2 in latency I type cell Rael (I) and latency III type cell CBMI Ral-STO (III). Monoclonal antibodies were used to detect the proteins as described in Methods. b) Chromatin immunoprecipitation(CHiP)-assay of EBNA-1 and Oct-2 probed by PCR over the left hand of FR. The antibody-protein cross-linked and precipitated DNA was probed undiluted, diluted 1:10 and 1:100. The sources of the protein-FR complexes were the same cell lines as in a). Controls were immunpreciptation leaving out the first specific antibody, a cloned, purfied piece of FR (positive control) and PCR with no DNA (negative control) c) Control of non-specific binding of antibodies to DNA. DNA from latency III cell lysate was immunopreciptated with anti-Oct2 or anti-EBNA1, with negative and positive controls. Results show no significant background binding of the antibody.