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

FIG.5.

FIG.5.

FIG.5.

(A to F) Inhibitory effect of Ubc9 on promoter transactivation by HHV-6A IE2. HEK293T cells were transfected as described in Materials and Methods with 1 μg reporter construct (pLTR-Luc encoding the HIV-1 LTR promoter or p2-1900 encoding the human COX-2 promoter), 0.1 μg transactivator (pcDNA4-IE2 or control pcDNA4-Tat), and 0, 0.12, 0.25, 0.5, or 1 μg of either pcDNA3.1-Ubc9 or pcDNA3.1-Ubc9 C93S. Cells were lysed 48 h after transfection. Transactivation of the reporters was quantified by measurement of luciferase activity expressed as transactivation (n-fold) relative to control (pcDNA-transfected cells). Results are means ± SD from three distinct experiments of duplicate transfections and were normalized for protein content in each sample. *, P < 0.05 compared with IE2- or Tat-transfected cells. (G) Effect of the deletion of the Ubc9-interacting domain of HHV-6A IE2 on HIV-1 LTR promoter transactivation. HEK293T cells were transfected with 1 μg pLTR-Luc reporter construct, 0.1 μg pcDNA4-IE2 or pcDNA4-IE2Δ993-1037, and 1 μg pcDNA3.1-Ubc9 or pcDNA3.1-Ubc9 C93S. Luciferase activity measurement is described in Materials and Methods. *, P < 0.05 compared with IE2-transfected cells.