Figure 5. TTP inhibits c-Jun transcription via zinc finger at 147.

MDA-MB-231 A. and MCF7 C. cells were infected with TTP/Ad and control adenovirus (EV/Ad) for different times as indicated, followed by extraction of total RNA to measure c-Jun primary transcript by real-time RT-PCR. The TTP expression in the above MDA-MB-231 B. and MCF7 D. cells were determined by real-time RT-PCR. c-Jun E. and TTP mRNA F. was measured by real time PCR in the TTP/Tet-Off MDA-MB-231 cells after adding Dox (2 μg/ml) for different times as indicated. Data represent are mean plus SEM of three experiments. 5 × 10⁶ HEK293 cells were transiently transfected with c-Jun promoter-driven luciferase construct along with different amounts of TTP expression vector, followed by measurement of luciferase activity in cell lysates G. 5 × 10⁶ HEK293 cells were transiently transfected with c-Jun promoter-driven luciferase construct along with TTP as well as two TTP zinc finger mutant constructs (C124R & C147R), followed by measurement of luciferase activity in cell lysates after 24 hours H. Data shown as relative levels compared to luciferase activity in cells transfected with the empty vector (pCMV). c-Jun protein levels were detected in the above cells by western blot with antibody against c-Jun protein I.