Validation of the novel method by analysis of core promoter elements.

<p>A) The analysis of basalactivator-independent transcription of the wild type E4 promoter and an E4 derivative that contains a defective BRE. B) The analysis of basal activator-independent transcription of the wild type ML promoter and an ML derivative that contains a defective BRE. C) The effect of TFIIB mutation (G153Q:R154A) on the activity of basal transcription for the E4 promoter (left panel) and ML promoter (right panel) using TFIIB-depleted and non—depleted NE supplemented with wild type TFIIB or its mutant. D) Analysis of transcription activation usingthe promoters BRE-mTATA (BREmT) and mBRE-mTATA (mBREmT), using nonradioactive <i>in vitro</i> transcription assay. E) As in part D but using a conventional <i>in vitro</i> transcription assay followed by electrophoresis and detection by autoradiography. F) The AdML promoter derivatives BRE-mTATA (BREmT) and mBRE-mTATA (mBREmT)linked to a luciferase reporter were co-transfected with a vector driving expression of the activator GAL4-VP16. 48 hours later the cells were lysed and luciferase activity was quantified. Each bar represents the mean of at least three independent experiments with standard deviation. The symbol “*” represents P≤0.05, the symbol “**” represents P≤0.01,the p values were obtained by performing <i>t t</i>est.