FIG 5.

HPV16 or HPV18 infection and viral E6 or E7 affect the expression of RBPs in raft cultures. Total tissue RNA from each raft culture was extracted and used for the study. (A to D) Expression of 8 RBP genes in 10-day-old raft tissue samples derived from human vaginal keratinocytes (HVK) (A [HVK and HVK16] and C [HVK and HVK18]) or human foreskin keratinocytes (HFK) (B [HFK and HFK16] and D [HFK and HFK18]) was validated by TaqMan RT-qPCR and compared to expression of raft cultures of HVK or HFK cell lines containing HPV16 (A and B) or HPV18 (C and D). (E to H) Validation by TaqMan RT-qPCR of RNASEH2A and NOVA1 expression in primary HFK-derived raft cultures with or without productive HPV18 infection or HPV18 oncogene expression. HPV18 genomes were excised from recombinant plasmid in primary HFK by Cre-LoxP recombination. (E and F) Raft cultures with or without productive HPV18 infection were prepared within a week of DNA transfection and followed by TaqMan RT-qPCR detection of the expression of RNASEH2A (E) and NOVA1 (F) in primary HFK-derived raft tissue samples on day 8 (D8), day 12 (D12), and day 16 (D16). (G and H) TaqMan RT-qPCR detection of the expression of RNASEH2A (G) and NOVA1 (H) in day 11 raft cultures derived from primary HFK acutely transduced with a retrovirus expressing HPV18 E6, E7, or E6E7 or with an empty control retrovirus. NS, not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001. The data in all panels represent means ± SD determined in duplicate from three independent raft tissue samples.