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. 2001 Apr;75(8):3841–3850. doi: 10.1128/JVI.75.8.3841-3850.2001

FIG. 2.

FIG. 2

FIG. 2

Accumulation of PV-specific RNAs in transfected cells. HeLa cell monolayers were transfected with RNA transcripts in the presence of DEAE-dextran. Cells were harvested at the indicated times after transfection. (A) Accumulation of plus-strand RNA. Total cytoplasmic RNA isolated from approximately 104 cells was bound to a nylon membrane and hybridized to a 32P-labeled riboprobe complementary to nt 5823 to 6056 of the vRNA. The standard curve shows increasing amounts of purified PV RNA (0.1 to 10 ng) hybridized in parallel. (B) Accumulation of negative-strand RNA. Total cytoplasmic RNA isolated from approximately 105 cells was subjected to two-step RNase protection (40). Positions of unprotected (258-nt) and protected (230-nt) probes are indicated. Top, mock-transfected cells (lanes 1 to 5) and cells transfected with PV full-length RNA transcript (lanes 6 to 10); bottom, cells transfected with PVΔP1 (lanes 1 to 5), E5PVΔP1 (lanes 6 to 10), and PVΔP1-3D∗ (lanes 11 to 15) RNA transcripts.