Figure 7. Acetylation modulates NS1 cytotoxic activities.

1. Mutation of the NS1 acetylation sites decreases H-1PV cytotoxicity. Cells were seeded into a 96-well E-plate and infected with mutant or wild-type (wt) viruses at the same MOI (10 Vg/cell). Cell proliferation was monitored in real time with the xCELLigence system as described in Fig 2D. The vertical grey line indicates the time of treatment.
2. Mutation of both NS1 acetylation sites decreases H-1PV-triggered intracellular ROS accumulation. HeLa cells were infected with 500 Vg/cell of H-1PV wild-type (wt) or H-1PV-NS1-K85R-K257R mutant virus. 24 h after infection, cells were loaded with DCFH-DA, harvested and analysed by FACS for ROS content as described in Fig 1B.
3. Mutation of the NS1 acetylation sites decreases H-1PV-triggered cell lysis. HeLa cells were infected with H-1PV wild-type (wt) or mutant derivatives (H-1PV-K85R, H-1PV-K257R, H-1PV-K85R-K257R) and grown for 72 h in the presence or absence of VPA before being processed for the LDH assay as described under Materials and Methods Section. Results shown are average cell lysis values with standard deviation bars, calculated from four replicates per experimental condition.