FIG. 3.

Rabies virus N phosphorylation affects both viral RNA transcription and replication in the minigenome system. BSR cells infected and transfected as described for Fig. 1 were harvested for RNA or protein analysis. mRNA was purified from total RNA, and genomic analogues were purified from immunoprecipitated RNP with polyclonal anti-N antibodies. Northern blots prepared from these two types of RNA preparations were hybridized with a CAT cDNA probe to assay mRNA transcripts (CAT mRNA) and genomic RNA (both genomic and antigenomic) analogues. [³⁵S]methionine- or [³²P]phosphoric acid-labeled N protein expressed in the cells was immunoprecipitated with polyclonal anti-N antibodies and analyzed by SDS-PAGE.