Fig 1: Inhibition of E2-activated SGK1 increases the phosphorylation and nuclear translocation of NF-?B in LPS-stimulated DSCs. Western blot analysis of LPS-stimulated DSC lysates pretreated with E2 in the presence or absence of GSK650394 (A-F). (A) Blots were probed for phosphorylated I?Ba (p-I?Ba), total I?Ba (t-I?Ba) and total ß-actin. (B) Densitometric quantifications of p-I?Ba and t-I?Ba to ß-actin (left), and mean (SEM) ratio of phosphorylated-to-total I?Ba protein (right). (C) Bolts were probed with phosphorylated NF?B (p-NF-?B), total NF-?B (t-NF-?B), and total ß-actin. (D) Densitometric quantifications of p-NF-?B and t-NF-?B (left) to ß-actin, and mean (SEM) ratio of phosphorylated-to-total NF-?B protein (right). Western blot analysis of cytosol (E) and nucleus (F) NF-?B p65 and ß-actin and nuclear TBP served as the loading controls. Western blot analysis of SGK1 (G), cytosol (H) and nucleus (I) NF-?B p65 of DSCs pretreated with SGK1-specific and non-targeting negative control siRNA. Control group, phenol red-free RPMI-1640 media; LPS group, 10 ng/mL LPS; LPS+E2 group, 10 ng/mL LPS + 10 nM E2; LPS+E2+GSK650394 group, 10 ng/mL LPS + 10 nM E2 + 10 µM GSK650394. Data are represented as arithmetic means ± SEM for 3 independent samples. **P < 0.01, ***P < 0.001, compared with control group or medium group; ??P < 0.01, ???P < 0.001, compared with LPS group or E2 group; ###P < 0.001, compared with LPS+E2 group or E2+control siRNA group.