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. 2006 Oct;80(20):10218–10228. doi: 10.1128/JVI.00375-06

FIG. 6.

FIG. 6.

Nuclear localization of IE2 in transfected cells is not altered by overexpression of Ubc9. HEK293T cells were transfected as described in Materials and Methods with 1 μg pLTR-Luc reporter construct, 2 μg pcDNA4-IE2 and 0, 1, 2, or 3 μg pcDNA3.1-Ubc9. Cells were harvested 48 h after transfection and split for the following assays. (A) Transactivation of the pLTR-Luc reporter was quantified by measurement of luciferase activity, expressed as transactivation (n-fold) relative to control (pcDNA-transfected cells). Results (±SD) represent one typical experiment out of three and were normalized for protein content in each sample. (B) The HEK293T-transfected cells were also processed for cellular fractionation into cytoplasmic (C) and nuclear (N) protein fractions as described in Materials and Methods. Samples were analyzed by SDS-PAGE and probed using anti-IE2 or anti-Ubc9 antibodies.