Figure 7. FBXL19 target genes rely on CDK-Mediator for activation during differentiation.

(A) A schematic illustrating the OHT treatment to generate MED13/13L KO ES cells and differentiation approach. (B) Western blot analysis showing the efficiency of MED13/13 knock-out upon 96 hr OHT treatment of Med13/13l^fl/fl ES cells. Suz12 was blotted as a loading control. (C) ChIP-qPCR showing CDK8 enrichment in WT and MED13/13L KO ES cells. Enrichment is relative to gene desert control region. Error bars show standard deviation of two biological replicates. (D) RT-qPCR gene expression analysis in WT and MED13/13L KO ES cells before and after RA induction. Expression was normalized to the expression of the PolIII-transcribed gene tRNA-Lys and is represented as relative to WT ES cells (for pluripotency genes) or RA-treated WT cells (for differentiation markers). Error bars show SEM of three biological replicates, asterisks represent statistical significance calculated by Student T-test: *p<0.05, **p<0.01, ***p<0.001.

Figure 7—figure supplement 1. FBXL19 target genes rely on CDK-Mediator for activation during differentiation.

A schematic representation of the strategy used to generate a MED13/13 l double conditional ERT2-Cre ES cell line. LoxP sites flanking exons 7 and 8 of Med13 and Med13l were inserted using CRISPR-Cas9 mediated targeting. Treatment with tamoxifen (OHT) results in deletion of exons 7 and 8.