Cytotoxic effects of transfected FasL constructs on Jurkat cells. Cytotoxicity is reflected by the percentage of Fas-positive Jurkat cells killed by co-culture with transfected 293T cells. (a) Ratios of 1:1 or 3:1 were used for 293T cells (effectors) transfected with either of 4 constructs (WT; Pt 86, black; Pt 55C, open; and hgld, shaded) to Jurkat cells (targets). Averages of % cell death and error bars are shown as normalized to that of WT FasL-induced death. Data represent at least 3 experiments. (b) Cotransfection of the Pt 86 (R156G) mutant FasL with wild type FasL. 293T cells were transfected with a total of 0.4 or 0.2 μg of wild-type FasL only (open bars); with Pt 86 mutant mixed with wild-type FasL at ratio of 1:1 (black bars); and with Pt 86 mutant FasL only (shaded bars). The ratio of effector to target cells was 2:1 in this experiment. Means and error bars of the percentage Jurkat cells death are shown. Data represent two independent duplicate experiments. (c) Surface staining of FLAG-tagged FasL in 293T cells transfected with empty vector, hgld-, WT-, and Pt 86- FasL as indicated.
Bi et al. BMC Medical Genetics 2007 8:41 doi:10.1186/1471-2350-8-41