Figure 3.

Explants respond to 1 µM progesterone (P4) in paired biopsies whereas myometrial cells do not. Myometrial tissue obtained from term non-labouring women was divided into three portions: (i) snap frozen (t = 0), (ii) finely dissected into 3 × 3 × 3 mm³ explants and (iii) digested with a collagenase mixture to isolate cells for myometrial cell culture. Explants were immediately pre-treated for 6 h with ethanol vehicle, 1 or 10 µM P4 followed by a 24 h treatment with IL-1β (10 ng/ml). After overnight incubation with 1% (v/v) charcoal and dextran-stripped fetal calf serum, myometrial cells at passage 4 were pre-treated for 6 h with ethanol vehicle, 1 or 10 µM P4 followed by a 24 h treatment with IL-1β (10 ng/ml). Protein was extracted and quantified. Western blotting for cyclooxygenase-2 (COX-2) was performed as described in Methods. A representative western blot is shown at the top of the figure with densitometric analysis below. The data are expressed as mean + SEM. Normality was tested using a Kolmogorov–Smirnov test followed by comparison of control versus IL-1β by the Wilcoxon-signed rank testing or paired t testing depending on the data distribution; *P < 0.05 versus control in that group. The IL-1β, 1 µM P4 & IL-1β and 10 µM P4 & IL-1β conditions in each group were compared by ANOVA followed by Bonferroni's Multiple Comparison Test; ^#P < 0.05 versus IL-1β in that group; ^##P < 0.01 versus IL-1β in that group, n = 4–8.