Fig. 4.

C7-equivalent homologs from diverse mammalian poxviruses bind SAMD9, and the binding is disrupted by mutations at the molecular claw. HeLa cells in six-well plate were infected with VACV expressing the indicated viral proteins at a multiplicity of infection (MOI) of 5 pfu per cell. All viral proteins had a C-terminal V5 epitope tag. VACV-A6, a viral protein involved in viral assembly, was used as a negative control. At 8 hpi, the cells were lysed. V5-tagged viral proteins were immunoprecipitated from the cell lysates with agarose conjugated with a monoclonal V5 antibody (IP with V5). The levels of SAMD9 and V5-tagged viral proteins in the cell lysates and precipitates were determined by Western blot with antibody against SAMD9 and V5, respectively. The experiment was performed twice with similar results, and a representative result is shown. Ig LC, the light chain of the anti-V5 antibody.