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. 2011 Mar 28;6(3):e18376. doi: 10.1371/journal.pone.0018376

Figure 6. KLK6 over expression in Jurkat T cells reduces cell death.

Figure 6

Jurkat T cells were stably transduced with a vector in which the human KLK6 gene is constitutively expressed under the control of a CMV promoter, or with an empty vector (Control), and levels of live (Annexin V-PE- and 7AAD−), early apoptotic (Annexin V-PE+ and 7AAD−), or dead (Annexin V-PE+ and 7AAD+) cells determined by flow cytometry under resting conditions, or after 24 hr periods of exposure to 0.1 or 0.25 µM staurosporine. (A) Histogram and corresponding dot plots (B), demonstrate KLK6 over expression produces effects largely parallel to those afforded by treatment of cultures with recombinant KLK6. There was a decrease in the percentage of dead cells and an increase in the number of live cells in Jurkat T cells over expressing KLK6, relative to those expressing an empty vector. KLK6 over expression also reduced cell death in the presence of 0.1 or 0.25 µM stuarosporine relative to that seen in cells stably transduced with empty vector. Reductions in cell death were likely to reflect in part a delay in apoptosis, since after 24 hr exposure to 0.25 µM Staurosporine, KLK6 over expression not only reduced the number of dead cells and increased the number of live cells, but cells in the early stages of apoptosis (AnnV-PE+, 7AAD−) were also significantly elevated. Data are expressed as mean ± SEM of triplicate cultures examined in parallel; One Way ANOVA with SNK post hoc test for multiple comparisons, P<0.001***, P = 0.002**, P = 0.003*. All data shown is representative of that seen in at least 3 independent cell culture experiments.