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. 2016 Jan 24;7(8):9069–9083. doi: 10.18632/oncotarget.6997

Figure 1. Transcription of MYB is attenuated at the pausing site within intron-I in breast cancer cells by CDK9i.

Figure 1

A. Schematic diagram of human c-MYB gene showing the promoter, intron-1 containing a stem-loop forming region followed by poly dT tract (SL-dT motif). Locations of primers used for the detection of intronic transcripts (Pre-I, Pre-II and Post-III, Post-IV), and for the mature transcript (exons 8 and 9) are indicated. B. CDK9i selectively downregulate the expression of MYB. MCF-7 cells were incubated with different CDK9i as shown (Flavo-, Flavopiridol; AT7519; BE-09-LN53;) for 4h. The concentrations of each drug in nM are shown next to the name of the drug. Expression of MYB was normalized using β-actin gene expression as an internal control. C. CDK9i induced pausing at the SL-dT region. MCF-7 cells were treated with the indicated CDK9i for 4h and qPCRs were performed using the indicated intronic primers. Relative expression of intronic transcripts measured by Pre- and Post- primer pairs was determined using a β-actin intronic transcript as an internal control. The degree of pausing is expressed as a ratio of relative expression of Pre to Post-transcription, normalised to untreated controls. All qPCRs (n ≥ 3) were performed in triplicate with error bars indicating standard deviation (SD).