Figure 6.

NR4A2 enhances the release of Pol II from the Prl promoter into the gene body. A. Schematic of primer locations used in subsequent assays. B. Pit-1/0 cells were transfected with a Pou1f1 expression plasmid or with a combination of expression plasmids encoding both Pou1f1 and Nr4a2 proteins. Transfection with an empty GFP vector served as a negative control. ChIP was performed on flow-sorted GFP+ cells with a Poll II antibody (Ser 5-P isoform), and occupancy was quantified by qPCR. The Gh, Pou1f1, and Prl promoters were assayed for Pol II occupancy along with regions 500 bp upstream and downstream of the respective promoters. * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001 as determined by 1-way analysis of variance. n.s. indicates no significance. n = 3 biological replicates. ChIP, chromatin immunoprecipitation; GFP, POL II, polymerase II.