One relates to your statement:After allowing the recombinant AT1-R mRNA to reach a steady-state, its transcription can be selectively suppressed by adding anhydrotetracycline (Antet). This approach reveals the instrinsic decay rate of the mRNA and avoids toxicities associated with the use of general transcriptional inhibitors such as actinomycin D, which might interfere with normal AT1-R mRNA regulation and many other cellular processes.
How does Antet inhibit transcription? It will be useful if you can guide me to a good reference on the mechanism of action, effectiveness of inhibition and selectivity of this agent.
The second relates to the stability of mRNA. I am impressed that you have PKA-dependent system. What mRNA domains are involved - coding region or anything in the UTR also?