DRB-PRO-Seq for measuring Pol II transcriptional elongation rate alteration up on CDK12 and CDK13 inhibition by THZ531

We performed PRO-Seq to measure the Pol II elongation rate by measruing the distance of Pol II travels at viarous time points after relased from DRB induced pausing. We compared DMSO/DRB or THZ531/DRB co-treated sample to assess the effect of CDK12 and CDK13 inhibtion by THZ531 on transcriptional elongation rate. Cells were treated a co-treatment cosistent of 100μM DRB and DMSO/100nM THZ531/200nM THZ531 for 4 hours, then DRB was washed out to allow Pol II to elongate for 12.5 mintues, 25 minutes and 45 minutes. Before wash-out, 0 minute time point were also haversted As THZ531 covelantly binds to CDK12 and CDK13, we can assess the effect of CDK12 and CDK13 inhibtion on elongation after DRB being washed out. Control samples were treated with DMSO/THZ531 100nM and 200nM for 4 hours but no DRB.