Fig. 6
E2 activates SGK1 and promotes the cytosol location of NF-κB in uterine macrophages of ovariectomized (OVX) mice. Eight to ten week old female ICR mice were divided randomly into a sham group and five experimental groups (n = 10). The mice in the five experimental groups were ovariectomized before treatment with different hormone combinations. A The uterine morphologies in mice injected with steroid hormones or ICI 182780. B Uterine macrophages were extracted from the uteruses of the female mice that received either vehicle (corn oil), E2, P4, E2 + P4, or the ER antagonist ICI 182780 after OVX. Final magnification: × 630, scale bar 25 μm. (C-G) Western blotting analysis in uterine macrophages of mice treated with E2 (100 ng) alone, P4 (1 mg) alone, E2 (100 ng) plus P4 (1 mg), ER antagonist ICI182780 (100 μg), or corn oil (100 μL) after OVX and in the sham group. C Blots were probed with antibodies to t-SGK1, p-SGK1, and β-actin. D Western blotting analysis of p-IκBα, t-IκBα, and total β-actin. E Blots probed for p-NF-κB, t-NF-κB, total β-actin. F Cytosol lysates of uterine macrophages probed for cytosolic NF-κB p65 and β-actin. G Nucleus lysates probed for nuclear NF-κB p65 and TBP in uterine macrophages. Data are the arithmetic means ± SEM for three biological replicates. **P < 0.01, ***P < 0.001, contrasted with the sham group; ∆∆P < 0.01, ∆∆∆P < 0.001, contrasted with the OVX control group; ##P < 0.01, ###P < 0.001, contrasted with the OVX + E2 group; + + P < 0.01, + + + P < 0.001, contrasted with the OVX + P4 group; &&&P < 0.001, contrasted with the OVX + E2 + P4 group. E2, estradiol; SGK1, serum-glucocorticoid regulated kinase; OVX, ovariectomized; P4, progesterone; ER, estrogen receptor; p-, phospho-; t-, total-; NF-κB; nuclear factor kappa B; IκBα, inhibitor of nuclear factor kappa-B kinase subunit alpha; TBP, TATA binding protein; SEM, Standard Error of the Mean