Figure 2.

HPV E2 does not act synergistically with HPV E1 or RF to stimulate pol ε. (A) Primed M13 assays were used to evaluate the combined effect of E2 with E1 or RF on pol ε. HPV 11 GST-E2 was added in increasing amounts (20 ng/μL to 200 ng/μL) to reactions containing pol ε (lanes 6–9), pol ε with 60 ng/μL of HPV E1 (lanes 12–15), and pol ε with RF (lanes 18–20). Lanes 3–5 and 10 represent controls. Lanes 11, 16, and 17 represent the stimulation of pol ε by either E1 or RF in the absence of E2. Lanes 1 and 2 show the same gel size markers as described in Figure 1. (B) Quantification of results of (A) and repeat experiments using phosphorimage analysis as described in the Methods. Pol ε + RF was set to 100% and all other values were plotted relative to this. Experiments were done three times and standard deviations were based on three experiments. An unpaired Student’s t test (GraphPad software) was used to compare the effect of E2 on pol ε with the combined effect of E2 and RF on pol ε. The p value was 0.26 and not found to be statistically significant.