Figure 2.

Progesterone stimulates PAD-catalyzed citrullination to regulate IGFBP1 mRNA expression. (A) OLE cells were stimulated with vehicle (DMSO) or 100 nM progesterone for 30 or 60 min. After cell lysis, histones were purified and equal amounts examined by Western blot. Membranes were probed with an anti-histone H3Cit 2, 8, 17 antibody or anti-total histone H3 as a loading control. The top panel shows a representative Western blot, while the bottom graph represents 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 = 3, *P < 0.05). (B) OLE cells were pretreated with vehicle (DMSO) or 2 μM BB-ClA for 3 h followed by stimulation with 100 nM P4 for 2 h. Total RNA was purified, reverse transcribed, and then cDNA was examined by qPCR with intron spanning primers for IGFBP1 and GAPDH as the reference gene control. All values are expressed as means ± s.e.m. Means were separated using SNK (n = 5, *P < 0.05).