FIG 5.

Tax fails to activate sense transcription in all cells. (A) Flow cytometry analysis of sense/antisense profiles of HEK293T cells cotransfected with the reporter construct and pcDNA3.1 (left plot) or pSG-Tax (right plot). Plots show one representative experiment from four independent experiments. (B) The graph shows mean cell percentages in each plot quadrant from the four independent experiments including those in panel A. Error bars denote standard deviations, and significant results as determined by Student's t test are indicated (**, P < 0.001). (C) The Western blot shows Tax expression in HEK293T cells transfected with the reporter construct and an empty vector (lane 1) or pSG-Tax-His (lanes 2, 3, and 4). The left panels show Tax and actin expression in unsorted cells. The right panel shows Tax and actin expression in cells sorted as a function of EGFP or EGFP/DsRed expression. (D) The graph shows quantitative real-time PCR amplification of DNA from cells transfected with pcDNA3.1 or pSG-Tax that were sorted as a function of their fluorescent signals. Values were normalized to those of the fluorescence-negative population (set to 1). The graph shows data averaged from three independent experiments. (E) Plots show separate analyses of antisense (EGFP) and sense (DsRed) transcription of the plots in panel A. (F) The graph shows mean cell percentages in each plot section from the four independent experiments. Error bars denote the standard deviations, and significant results as determined by Student's t test are indicated (*, P < 0.01; **, P < 0.001).