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. 2006 Jul;80(14):7052–7059. doi: 10.1128/JVI.00103-06

FIG. 1.

FIG. 1.

TORC proteins are transcriptional coactivators of HTLV-1 Tax. (A) Expression of TORCs in HeLa cells. Cells were transfected with Tax expression plasmid pIEX alone (lanes 1 and 5) or with pIEX plus pcDNA3.1-V5-TORC1/2/3 (lanes 2 to 4 and 6 to 8). Cell lysates were analyzed by Western blotting with rabbit anti-V5 (Invitrogen) or mouse anti-Tax (11). The expression of TORCs did not affect the activity of the cytomegalovirus promoter. (B to D) Cooperation of TORCs and Tax in the activation of HTLV-1 LTR. HeLa cells (2 × 105 per well) were transfected with reporter plasmid pLTR-Luc (1 μg) plus the indicated combinations of expression plasmids. A fixed amount of Tax expression plasmid pIEX (500 ng) and escalating amounts of expression plasmids (60 to 250 ng) for TORCs were used. (E to G) Cooperation of TORCs and Tax in the activation of TRE. HeLa cells (2 × 105 per well) were transfected with reporter plasmid pTRE-Luc (1 μg) plus the indicated combinations of expression plasmids. A fixed amount of Tax expression plasmid pIEX (500 ng) and escalating amounts of expression plasmids (60 to 250 ng) for TORCs were used. (H to J) Stimulation of Gal-Tax-mediated transcriptional activation by TORCs. HeLa cells (2 × 105 per well) were cotransfected with reporter plasmid pGal-Luc (1 μg) and expression plasmids for Gal-Tax (500 ng) and TORCs (60 to 250 ng). Cells were harvested 48 h after transfection, and relative luciferase activities (RLA) in arbitrary units were calculated from the readouts of firefly luciferase normalized to the readouts of Renilla luciferase. The results represent three independent experiments, and the error bars indicate standard deviations.