Fig. 4

Temporal expression, localization, and interactions of replisome core proteins. A Western blot analysis of vHelicase/primase (ADRV-88L) and vPCNA (ADRV-23L). GSTC cells were infected by ADRV at 0.5 MOI and collected at indicated time points. B Localization of ADRV-88L by immunofluorescence in GSTC cells. ADRV-88L (green) was stained with an anti-88L antibody, and vSSB (red) was stained with an anti-vSSB antibody. Representative overlaps are indicated by white arrows. C Localization of ADRV-23L by immunofluorescence. ADRV-23L (green) was stained with an anti-23L antibody. Representative overlaps are indicated by white arrows. Visible Hoechst-labeled cytoplasmic viral factories are indicated with yellow arrows. D Interactions among the four replisome core proteins of ADRV (23L, 47L, 85L, and 88L) revealed by coimmunoprecipitations (co-IP). Cell lysates from HEK293T cells cotransfected with plasmids expressing the indicated proteins and immunoprecipitated (IP) protein complexes were subjected to Western blotting (WB) analysis using anti-HA or anti-Flag antibody. The interactions between two proteins are marked with black lines. A protein pair without interaction is indicated by a broken line