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G mutants induce death m several tumor celi lines.

To investigate whether the strong inhibition of cellular transcription and translation observed for all G mutants would provide them with increased oncolytic properties, we assessed their ability to kill various murine and human tumor celi lines in vitro (Figurę 5). Their capacity to induce death of the murine 3LL pulmonary carcinoma (Figurę 5B), the murine EL4 and human Jurkat lymphomas or the human HepG2 hepatocellular carcinoma (data not shown) was comparable to WT VSV or the M mutant. Importantly however, all G mutants were morę efficient at killing murine B16 melanoma cells than the M mutant (P < 0.001 at 48h; P < 0.01 at 60h), while Gsr, Gó and Gór also showed a significantly faster induction of B16 celi death compared to wild-type VSV (Gsr and Gór compared to WT P < 0.01 at 24h; Gó compared to WT P < 0.05 at 24h) (Figurę 5C). In addition, Gsr and Gó showed an increased kilhng of MC57 fibrosarcoma cells at 48h post infection compared to WT VSV (P - 0.01 for Gsr ; P < 0.05 for Gó) while Gsr, Gó and Gór also induced a significantly higher death compared to the M mutant (P < 0.01) (Figurę 5D). As a control, we analyzed celi death in L929 fibroblasts and showed that G mutants induce significantly slower death than the M mutant on these cells (P < 0.01 for Gs, Gsr and Gó and P < 0.05 for Gór at 24h; P < 0.001 for Gs, Gsr and Gó at 36h) (Figurę 5A). Finally, Gsr and Gór also induced an increased death of L929 cells compared to WT (P < 0.0001 for Gór and P < 0.05 for Gsr). We conflrmed these results using a trypan blue exclusion assay that showed the same trend for L929 and B16 celi lines (data not shown).