Figure 5.

Recruitment of P-TEFb from storage complexes into AF4/AF4N. A. Schematic representation of the 7SK RNP. B. Western blots after CDK9 immunoprecipitation from whole cell lysates of transiently transfected 293T cells (mock, Strep-tagged AF4, HCMV IE1 or 5-LO). blots were probed with antibodies against CDK9 and LARP7. C. 7SK RNA binding assay. Results of specific quantitative PCR of 7SK RNA purified from elution fractions. Samples were subjected to quantitative PCR and quantified against a 7SK RNA dilution series. Shown is the x-fold binding of 7SK RNA compared to the GST negative control (set as 1) [n = 3, +S.E.M., *p < 0.05]. D. Results of quantitative PCR of 7SK RNA after incubation of whole cell lysates with RNase, recombinant GST-AF4N or a distinct whole cell lysate with overexpressed AF4ST. A non-treated lysate was used as negative control. Shown is the relative amount of 7SK RNA after incubation as the quotient of 7SK RNA/β-Actin copy number.