Figure 5.

Progesterone stimulates calcium influx via L-type calcium channels to mediate PAD2 nuclear translocation. (A) OLE cells were pre-treated with vehicle (DMSO) or 10 nM nicardipine for 30 min then stimulated with vehicle (DMSO) or 100 nM P4 for an additional 30 min. Following cell lysis, chromatin-associated nuclear proteins were isolated and equal amounts were examined by Western blot. Membranes were probed with an anti-PAD2 antibody or anti-total histone H3 as a loading control. The left panel shows a representative Western blot, while the graph on the right illustrates the quantification of multiple Western blots using BioRad Image Lab 4.0. Data are presented as means ± s.e.m. and separated using SNK (n = 4, *P < 0.05). (B) OLE cells were grown on glass bottom dishes overnight then fixed, permeabilized, and examined by immunocytochemistry using an anti-PAD2 antibody (green) and stained with DAPI (blue). Cells were imaged using a Zeiss LSM 980 confocal microscope using a 63× objective and scale bar is 10 µm.