Fig. 5

The Mediator complex regulates response to CDK9i. Genome-wide loss of function CRISPR library screening was carried out in U-2932 and SU-DHL-10 cell lines as described in the methods. Data was analyzed using the MaGeCK pipeline. A Volcano plot of library screen data in SU-DHL-10 and U-2932 cells. Dots represent the log2(mid fold change) vs. –log10(mid p-value) of all sgRNA for one gene in CDK9i-treated cells versus control. Genes with a fold change significance of p < 0.1 are depicted in blue and select genes are highlighted in red and identified with a label. B-C Gene set enrichment analysis of the library screening was carried out with WebGestalt software. B Enrichment plots from U-2932 cells using the Cellular Component gene ontology. C Significantly enriched gene sets in SU-DHL-10 and U2932 cells are shown as bar graphs using the Cellular Component gene ontology. D MED12 knockout was established in U-2932 and VAL cells using RNP electroporation as described in the methods. Whole cell lysates were subjected to immunoblotting. Cells were treated with AZD4573 or vehicle control at the indicated concentrations for 48 h. Proliferation was quantified using a colorimetric tetrazolium-based assay. Mean ± SEM is shown. *p < 0.05 and **p < 0.01 vs. NT control. A table of IC50 values is included to the right