Research Paper Volume 1, Issue 9 pp 803—817

Figure 7. E6-AP 3' UTR is necessary for TTP-mediated decay. (A) HeLa Tet-Off/TTP-Flag cells were transfected with an expression vector containing the coding region of E6-AP (E6-APΔ3'UTR). Cells were grown in the absence or presence of TTP for 48 hr and lysates were analyzed for E6-AP and TTP protein expression by western blot. Actin was detected as a loading control. (B) HeLa cells were transfected with a luciferase-reporter construct containing the 1.6 kb E6-AP 3'UTR (Luc+E6-AP 3'UTR) or the control luciferase vector (no 3'UTR) along with a TTP expression construct (pcDNA3-TTP-Flag) or empty vector. Relative luciferase reporter activity in the absence (black bars) or presence (grey bars) of TTP is shown. Relative activity was assessed as luciferase activity normalized to its respective protein concentration for each transfection in the absence or presence of TTP. The data shown is the average of duplicate experiments. (*) P < 0.01