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Figure 2 | Retrovirology

Figure 2

From: A single point mutation in cyclin T1 eliminates binding to Hexim1, Cdk9 and RNA but not to AFF4 and enforces repression of HIV transcription

Figure 2

HA-CycT1-V107E mutant inhibits HIV transcription in human cells. a) HA-CycT1-V107E inhibits HIV-Tat dependent transcription from the HIV LTR promoter - HEK-293T cells were transfected with increasing concentrations of the HA-CycT1-V107E mutant and the LTR-Tat-BFP provirus lentivector using Lipofectamin 2000. 48 hr. post transfection cells were harvested and the percentage of cells that expressed BFP was measured by FACS. Data are presented relative to cells that did not express HA-CycT1-V107E - set to 100. Lower panel shows Western blot that confirmed HA-CycT1-V107E expression and equal expression levels of actin. b) HA-CycT1-V107E does not inhibit basal transcription from the CMV promoter - HEK-293T were co-transfected with HA-CycT1-wild type or HA-CycT1-V107E mutant in the presence of the CMV-BFP reporter promoter. 72 hr. post transfection, cells were harvested and their BFP expression was monitored by FACS. The percentage of BFP expressing cells in the presence of HA-CycT1 wild type was set to 100. Results are the average of there independent experiments. Error bars show ± SEM values.

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