Fig. 2.

Ongoing transcription but not translation is required for the induction of GR targets. U2OS-rGR cells were either pretreated for 1 h with 5 μg/ml actinomycin D, after which dex was added to 100 nM as indicated and incubation continued for 2 h (A); or treated with 100 nM dex for 2 h in the absence or presence of 10 μg/ml cycloheximide (B). Total RNA was harvested, reverse-transcribed, and subjected to real-time PCR with primer pairs to indicated genes. The data were transformed by the standard ∂∂Ct method, with amplification of ribosomal Rpl19 RNA used as an internal control for normalization.