Fig. 7

HSV-1 restriction by MxB is specified by its amino terminus and requires GTPase function. a HeLa cells were transiently transfected with pGL-T9G reporter plasmid together with increasing amounts of expression plasmids encoding the indicated proteins. At 24 h post transfection, cells were mock-infected or infected with HSV-1 strain MacIntyre (MOI = 0.5). Virus infection efficiency was determined by luciferase assay at 32 h post infection and is presented relative to the respective conditions with GST expression. Luciferase activity from all conditions is normalised to mock-infected cells. Data are representative of three independent experiments. Bars indicate mean ± s.d., n = 3 biological replicates. ANOVA modelling relative reporter activity with protein variant and dose as explanatory variables: protein variant, F(4,30) = 51.311, p < 0.0001; dose, F(2,30) = 29.844, p < 0.0001; synergistic interaction, F(8,30) = 6.407, p < 0.0001. Multiple comparisons: NS p ≥ 0.05; ****p < 0.0001. GST and MxB protein expression was controlled by immunoblot analysis using pooled samples for each condition. GAPDH served as loading control. b HeLa cells were transiently transfected with expression plasmids encoding the indicated proteins. At 24 h post transfection, cells were fixed and MxB protein expression and intracellular localisation was assessed by immunostaining. Nuclei were stained with Hoechst 33342. Scale bar, 20 µm