FIG. 5.
Steady-state, intracellular distribution of pPR, pAP, MCP, and other capsid and tegument (Teg.) proteins in cells infected with WT virus or the UL80 NLS mutants. HFF cells in 6-well plates were infected with WT, NLS1−, or NLS2− virus, scraped into the growth medium, collected by microcentrifugation at 4°C, and separated into NP-40 cytoplasmic and nuclear fractions as described in Materials and Methods for cells from 24-well plates. The samples were subjected to SDS-PAGE and analyzed by Western immunoassay as described in Results and Materials and Methods. Shown are images of the resulting membranes probed with antibodies as follows. (A) The UL80 proteins, including pPR and pAP and their cleavage products (PR and AP), were detected with anti-pAPhr; (B) MCP and a 120-kDa fragment (asterisk) were detected with anti-MCP; (C) mCP was detected with anti-mCP; (D) mC-BP was detected with anti-mC-BP; (E) SCP was detected with anti-SCP; (F) HMWP was detected with anti-pUL48c; (G) BPP/pp150 was detected with anti-BPP; and (H) LM/pp65 was detected with anti-LM. (I) Shown are the percentages of each protein in the NP-40 cytoplasmic (Cyto) and NP-40 nuclear (Nuc) fractions of the infected cells, calculated from the phosphorimages shown in the panels A to H.