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. 2001 Oct;75(19):9059–9067. doi: 10.1128/JVI.75.19.9059-9067.2001

FIG. 4.

FIG. 4

Specific binding of M protein to non-MHV RNA transcripts containing the packaging signal. (A) Schematic diagrams of the structures of plasmids PS5A and PS5B190. T7 Pr, T7 promoter; T7 Ter; T7 terminator. (B) Northern blot analysis of expressed RNA transcripts in RNA-expressing, MHV-infected cells (intracellular [i.c.] RNA) and packaged RNA transcripts in MHV particles (virion RNAs). PS5A RNA transcripts or PS5B190 RNA transcripts were independently expressed in MHV-infected cells. Intracellular RNAs and virion RNAs were analyzed on Northern blots using a probe that binds to the CAT sequence. The arrows indicate expressed RNA transcripts. (C) Specific binding of M protein to the expressed PS5B190 RNA transcripts in MHV-infected cells. Cell lysates were prepared from MHV-infected cells expressing non-MHV RNA transcripts at 12 h post-MHV infection. Anti-N protein monoclonal antibody (anti-N) and anti-M protein monoclonal antibody (anti-M) were independently added to equal volumes of cell lysates, and immunoprecipitation was performed. RNA was extracted from the immunoprecipitated samples. Intracellular (i.c.) RNAs and coimmunoprecipitated RNAs were analyzed using Northern blot analysis with a probe that binds to the CAT sequence. The arrows indicate expressed RNA transcripts.