FIG. 2.

RT-PCR analysis of HPV31b transcripts following infection of various cell lines. Subconfluent cell monolayers were inoculated with serial 10-fold dilutions of an HPV31b stock. Total RNAs were extracted and treated with DNase I. RT was preformed on 3 μg of RNA for each sample. Each cell line was mock-infected (lanes M), or infected with HPV31b doses corresponding to 0.1, 1.0, and 10 vDNA-containing particles per cell. CIN-612 9E monolayers (9E) and no RNA input (Ø) served as positive and negative amplification controls, respectively. (A) Primers E7.3A and E4.3B target a 498-bp amplimer arising from spliced HPV31b E1∧E4 RNA. The input RNA corresponded to 2.7 μg for each PCR. (B) Primers β-actin OA and β-actin OB detect a 641-bp amplimer derived from spliced cellular β-actin RNA. The input RNA corresponded to 0.3 μg. Molecular size standards (100-bp ladder, New England Biolabs) are shown at the edges of each panel.