Figure 2.

The effects of protein- and RNA-DE on effective reporter half-life. Tet-off HeLa cells were transiently transfected with TRE-driven reporter plasmids encoding Rn (A); Luc (B); HcRed (C); EGFP (D); ECFP (E); or EYFP (F). Reporter activity was measured following the inhibition of transcription by Dox treatment and expressed as a percentage of t = 0 h values. Results are expressed as means ± SEM (n = 4) of a single experiment (A and B) or are representative of at least 3 similar experiments (C–F); and show standard reporter constructs (open square) as well as those containing; the P1 protein DE (closed diamond), P2 protein DE (closed square), N4 RNA DE (open circle) or both P1 protein DE and N4 RNA DE (closed triangle). In the case of EYFP (F), an additional construct that further contained an N-terminal cUb was included. The results show a substantial reduction in effective half-life when both protein- and RNA-DE are included in the reporter construct.