Fig. 6

Functional analysis of host TopIIα and TopIIβ in ranavirus infection. A Cell viability of GSTC after doxorubicin treatment for 24 h. B Observation of virus infection after doxorubicin treatment. GSTC cells treated with doxorubicin were infected with recombinant ADRV expressing mCherry and observed at 24 hpi. C Determination of virus titers after drug treatment. The virus titers were determined by TCID₅₀ methods, which showed that ADRV titers were reduced by drug treatment. D Detecting viral relative genome copy numbers by qPCR. The relative genome copy numbers were determined by calculating the relative MCP copy numbers. Significant differences (versus virus without drug) are marked with *(p < 0.05). E Subcellular localization of host TopIIβ in ADRV-infected GSTC cells by immunofluorescence. Cytoplasmic TopIIβ (red) was colocalized with EdU labeling (green). Representative merges were indicated by white arrows. F Detection of the interaction between TopIIα and ADRV-23L by co-IP. G. Detection of the interaction between TopIIβ and ADRV-88L by co-IP. Cell lysates from HEK293T cells cotransfected with plasmids expressing the indicated proteins and immunoprecipitated (IP) protein complexes were subjected to Western blot (WB) analysis using anti-HA or anti-Flag antibody