Figure 2

A) Inhibition of GFP reporter translation mediated by expression of protein toxins. HeLa cells were transiently co-transfected with indicated toxin cDNAs and a GFP reporter. Inhibition of GFP translation, relative to pcDNA3/GFP co-transfected cells, as determined by fluorimetric analysis of cell lysates. B) Exogenous expression of α-sarcin, α-sarcin-mychis, α-sarcin-R121Q, and α-sarcin-R121Q-mychis in HeLa cells. Anti-sarcin western blot analysis of HeLa cell lysates transfected pcDNA3 or α-sarcin cDNA. * indicates the position of α-sarcin. R121Q mutant migrates slightly faster than wildtype α-sarcin. C) HeLa cells were transfected with either empty vector (MSCV) or 3 × Flag-sarcin. Stable cell lines expressing Flag-tagged-sarcin were established. Anti-Flag western blot of MSCV or 3 × Flag-sarcin transfected cells. Charted data represents the compiled results of 3 independent experiments, each performed in triplicate, +/- standard deviation.