FIG 2.

Rad51 and BRCA1 promoter activity and transcript levels are increased in HPV31-positive cells. (A) HFKs and HPV31-positive CIN612 cells were transiently transfected with luciferase reporter constructs containing either Rad51 or BRCA1 proximal promoter sequences, along with a Renilla luciferase-expressing control vector. Luciferase assays were performed 72 h posttransfection. Luciferase activity was normalized to Renilla luciferase activity, and values were calculated as fold change over luciferase activity in uninfected HFKs. Values are averages from five independent experiments ± standard errors of the means. *, P ≤ 0.03. (B and C) RNA was extracted from uninfected HFKs (B and C), HPV31-positive CIN612 cells (B), and HFK-31 cells (C). Quantitative reverse transcription-PCR analysis was performed using primers specific to Rad51 and BRCA1. Fold change was calculated using the 2^−ΔΔCT method. Shown is the fold change in message levels relative to that in uninfected HFKs, which is set to 1. Values are averages from five independent experiments ± standard errors of the means. *, P = 0.003; **, P = 0.0003. (D) Quantitative reverse transcription-PCR was used to examine transcript levels of Rad51 and BRCA1 in primary HFKs and HFKs that stably express HPV31 E6, E7, or E6/E7. Expression levels are shown relative to uninfected HFKs and were calculated using GAPDH as the reference gene. Shown is the relative fold change in gene expression over seven independent experiments, across three HFK backgrounds. Data are means ± standard errors. *, P ≤ 0.02; **, P ≤ 0.01.