FIG. 4.

Expression of individual MVC proteins or in combination by transfection does not induce cell death or cell cycle arrest. WRD cells were transfected with various constructs, as shown, that express GFP-fused MVC proteins. (A) Transfected cells were costained with annexin V-PI at 48 h posttransfection. The GFP-positive population was selectively gated. The percentage of nonapoptotic cells (annexin V⁻/PI⁻ population) in the GFP-positive population in each transfection was then plotted. All the values were generated from three independent experiments and are shown as an average with a standard deviation. (B) Representative results of cell cycle analysis. Transfected cells were stained with DAPI at 48 h posttransfection. The GFP-positive population was selectively gated. The cell cycle analysis results are shown as GFP-negative population versus GFP-positive population. (C)The percentage of cells at the G₀/G₁ phase is shown as an average with a standard deviation obtained from three independent experiments. (D) Transfected cells were stained with DDAO at 48 h posttransfection. The GFP-positive population was selectively gated and plotted. Numbers as shown are percentages of proliferated cells. The line as shown is arbitrarily set in the control cells, which were fixed immediately after transfection.