Figure 2

Kinetics of activator and reporter induction (A) and activation of an independent reporter in pCG6.0WCS/T (B-J). A; Transcriptional induction of the activator Gal4/Vp16 and the internal reporter (CFP) in pCG6.0WCS/T was analyzed by RT-PCR. Embryos were heat-shock for 90 seconds at st20/31.5hpf and were allowed to recover for the indicated periods of time. Activator transcripts were detectable already 10 minutes after induction and the levels increased up to 3 hours. Degradation to undetectable levels was complete after 20 hours. Activator dependent transcription of the internal reporter CFP was observed only after 2 hours and levels were still increasing after 25 hours. C-actin was used as an internal control. (B-J); The independent reporter plasmid pCG3.0Y was injected into the transgenic Gal4/Vp16 activator line. Anterior is to the left (B-J). Expressions of both internal and independent reporters were observed in a weak to strong ubiquitous manner (B-J). Occasional higher levels of internal reporter expression were observed in some parts of the embryonic body. These higher levels were paralleled by independent reporter expression (B-D, H-J). Mosaic clones exerting stronger YFP expression reflect locally higher plasmid concentrations (B-J). Abbreviations: h, hours; hpf, hours post fertilization; HS, heat-shock; M, size marker; st, developmental stage.