Fig. 4. NuMA and its relation to viral proteins and replication compartments. (A-C) HEp-2 cells were infected with 14R-VP16G at a MOI of 1 PFU/cell. Cells were fixed at 8 hours post infection and analysed for VP16-GFP and NuMA by indirect immunofluorescence. VP16-GFP (A) and NuMA (B) localised almost exclusively of each other. Note the difference in NuMA localisation between an uninfected cell (arrowhead) and an infected cell (arrow) (B). (D-G) Further analysis with triple fluorescence of VP16-GFP (D), UL17 (a tegument protein) (E) and NuMA (F) showed that, whereas the two viral proteins colocalised in the nucleus, NuMA localisation excluded these proteins. Analysis of NuMA and ICP8 localisation in mock-infected (H) and infected cells (I-K). Cells were infected with HSV-1 at a MOI of 3 PFU/cell and then fixed at 12 hours post infection. NuMA and ICP8 (a marker for replication compartments) exhibited mutually exclusive localisation.