A poxvirus pseudokinase represses viral DNA replication via a pathway antagonized by its paralog kinase
Fig 6
B12 exhibits a nuclear localization in uninfected and infected cells.
(A) CV1 cells with or without HA-B12 (GenScript) mRNA transfection were used for immunofluorescence detection of HA-tagged B12 (red, top row). CV1 control and CV1-B1myc expressing cells were incubated with αmyc for B1myc detection (red, bottom row). All cells were stained with DAPI nuclear stain (blue). (B) B1myc expressing CV1 cells were also transfected with HA-B12 (GenScript) mRNA and separately incubated with a primary antibody to detect HA-tagged B12 (αHA, top red image) or myc-tagged B1 (αmyc, bottom red image) and DAPI (blue) nuclear stain. (C) CV1 cells were infected with WT or WT/HA-B12 virus at a MOI of 5 and fixed at 4hpi or (D) 7hpi for immunofluorescence analysis of HA-B12 detection (red), I3 ssDNA binding protein (green) and DAPI nuclear stain (blue). The scale bars represent 100μm.