The F12/KLC2 interaction requires expression of E2.

<p>SDS-PAGE and immunoblot analysis of F12/KLC2 co-immunoprecipitations carried out in the presence of infection with a panel mutant VACVs lacking individual IEV proteins. Clarified cell lysates were generated from HEK TRex-F12-HAco cells that had been transfected with pFlag-KLC2 and 24 h later infected with VACV WR (vvWR), vΔF12, vΔA33, vΔA34, vΔA36, vΔB5, vΔF13, vΔE2 or mock-infected. These cells had also been induced to express F12-HA by addition of doxycycline (+Dox). One sample was transfected with pFlag-GFP as a negative control to measure background levels of co-immunoprecipitated F12-HA. (A) Input cell lysate samples were immunoblotted for Flag (i, KLC), HA (ii, F12), tubulin (iii, loading control) and D8 (iv, infection level control). Cell lysates were subjected to anti-Flag IP (B) and anti-HA IP (C), and co-precipitating proteins were analysed by blotting for Flag (i) and HA (ii). The positions of molecular mass standards (kDa) are shown on the left.