Figure 6.

ChIP of POU1F1 in Pit-1/Triple cells expressing Nr4a2 vs Pit-1/Triple cells that do not express Nr4a2. Pit-1/Triple cells, which do not express Nr4a2, were transfected with the same Nr4a2-IRES-GFP plasmid used in assays presented in Fig. 4C, and control cells were transfected with the empty IRES-GFP vector. GFP⁺ cells were sorted by FACS, chromatin was isolated, and ChIP was performed using an antibody that recognizes POU1F1. Following immunoprecipitation, qRT-PCR was used to measure the relative levels of POU1F1 binding in Pit-1/Triple cells that had been transfected with Nr4a2 plasmid (+Nr4a2), as well as those transfected with empty vector. Immunoprecipitations using IgG served as a negative control. *P < 0.05.