FIG. 4.

Effects of nucleosome assembly proteins on transcriptional activation by EBNA1. (A) Western blots of extracts of CNE2Z cells after transfection with siRNA against GFP (negative control), NAP1, NAP2, or TAF-I. Duplicate samples are shown. (B) After transfection with the indicated siRNA (or with siGFP in the second columns), cells were transfected with an EBNA1 expression plasmid or an empty plasmid (first column in each histogram), an FR-CAT reporter plasmid that is EBNA1 dependent, and a SEAP reporter plasmid that is independent of EBNA1. Effects on CAT expression (left) and SEAP expression (middle) were determined separately. CAT levels were also normalized to SEAP levels to account for any nonspecific transcriptional effects (right). **, P < 0.001 relative to the EBNA1 positive control. (C) CNE2Z cells were transfected with a plasmid overexpressing NAP1, NAP2, TAF-Iα, or TAF-Iβ or with empty plasmid (second columns) prior to transfection with EBNA1 expression, CAT reporter, and SEAP reporter plasmids and calculation of transcriptional activities as in panel B.