My drug is increasing proliferation of gene of interest (GOI). I would like to demonstrate the effect of this drug at the transcriptional level.
Im planning to check if it delays mRNA degradation of my GOI. Has anyone used actinomycin D and correct if Im wrong in how to use it:
1. treat cells with actinomycin D for various time points
2. measure mRNA of GOI by qPCR
3. Also detect GOI by protein
4. To ensure actiomycin D is working, I check cyclin D1 mRNA levels after actiomycin D treatment?