Figure 4

Effects of HPV 3' early regions on the steady state level of GUS mRNA and GUS protein. a) Schematic representation of the HPV sequences that were cloned into the retroviral HRSp-GUS reporter vector [26,27]. b) Northern Blot analysis of mRNA from SiHa cell pools (only GUS probe 1 experiments shown). c) GUS mRNA steady state levels. For each construct the steady state levels of the GUS mRNA and PURO mRNA were determined with real-time RT-PCR. The GUS mRNA levels were normalized with the PURO mRNA. The data plotted is the average of results from three independent pools of infected SiHa cells expressed relative to the empty HRSp-GUS construct. The error bars indicate the standard error of the mean. d) Protein produced per reporter mRNA For each construct the steady state GUS activity and total protein levels were determined. The GUS activities were normalized with the total protein levels. The protein/RNA ratios of the different reporter mRNA was evaluated by calculation of the ratio between the steady state levels of GUS protein to GUS mRNAs (not normalized for PURO). The data plotted is the average of results from three independent pools of infected SiHa cells expressed relative to the empty HRSp-GUS construct. The error bars indicate the standard error of the mean.