Figure 3.

Role of CRH on IL-1β-induced p65 phosphorylation at Ser536 in term nonlaboring myometrial cells. Primary cells were isolated from myometrial tissues obtained from pregnant women undergoing elective cesarean section for nonmaternal problems at term before the onset of labor. Cells were treated with CRH (100 nM) or IL-1β (1 ng/mL) or both for various time intervals (15 or 120 minutes); p65 S536 phosphorylation was monitored in nuclear or cytosolic fractions (A) or total cell extracts (B) by immunoblotting using specific antibodies as described in Materials and Methods. Total p65 was detected by immunoblotting. The nuclear protein TBP-protein was used as a loading control, and β-tubulin was used as a marker of cytoplasmic protein contamination. Representative Western blots are shown. The data represent the mean ± SEM of 3 estimations from at least 5 patients. *, P < .05 compared with basal (untreated); +, P < .05 compared with IL-1β alone.