A, LβT2 cells were plated on glass bottom confocal dishes, serum starved for 6 hours, and then treated with either vehicle or 10nM GnRHa for 0, 30, or 60 minutes. Cells were then fixed in 4% PFA and probed with an anti-PAD2 antibody with the appropriate fluorescently labeled secondary antibody and stained with DAPI. Cells were imaged with a confocal microscope using a ×40 objective. B, LβT2 cells were serum starved for 6 hours, then treated with either vehicle or 10nM GnRHa for 0, 30, or 60 minutes. The chromatin-associated protein fraction was isolated and quantified, and equal concentrations were examined by Western blotting. Membranes were probed with an anti-PAD2 and with antihistone H3 total as a loading control.