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. 2006 Sep 11;103(38):14188–14193. doi: 10.1073/pnas.0605985103

Fig. 3.

Fig. 3.

EBNA1 lacking UR1 can support extrachromosomal DNA replication and latent gene expression. B cell clones obtained with 2089 wild-type EBV, 3168 (ΔUR1-EBNA1), or 3169 (ΔUR2-EBNA1) from limiting dilution assays were expanded and analyzed for genomic extrachromosomal DNA molecules and latent gene expression. (A) Extrachromosomal viral DNA was detected by Gardella gel hybridization. The autoradiogram of the blotted Gardella gel hybridized with a specific, 32P-labeled plasmid probe revealed signals typical of extrachromosomal copies of EBV, as indicated by arrows (29, 30). Signals from 1 × 106 cells per lane with clones infected with 2089 wild-type or 3169 (ΔUR2-EBNA1) gave signals similar to Raji cells, which contain ≈50 copies of EBV per cell as a reference. Clones obtained with 3168 (ΔUR1-EBNA1) had a lower copy number, as indicated by weaker signals, with one case (3168.I) not visible in this exposure of the autoradiogram. (B) The expression of latent EBV proteins such as EBNA1, EBNA2, EBNA3A and -C, and LMP1 is similar among clones of B cells established with wild-type EBV and EBNA1 mutant viruses, as detected by Western blot analysis.