Figure 5.

NR4A2 occupancy at the Prl promoter is POU1F1 dependent. Pit-1/0 cells were transfected with a plasmid expressing Nr4a2 or with plasmids expressing both Nr4a2 and Pou1f1. All expression vectors contained an IRES/GFP reporter. GFP+ transfected cells were flow sorted, and chromatin was isolated for NR4A2 ChIP analysis. NR4A2 occupancy was assayed at the promoters of Gh, Prl, Pou1f1, and Myod genes (MyoD served as a negative control). * indicates P < 0.05 as determined by 1-way analysis of variance (n = 3 biological replicates). ChIP, chromatin immunoprecipitation; GFP, green fluorescent protein; IRES, internal ribosome entry sequence; NR4A2, nuclear orphan receptor transcription factor; POU1F1, pituitary-specific master regulatory transcription factor.