Fig. 4
SGK1 inhibition blocks E2-triggered M2 macrophage transition and Th2 immune responses in THP-1-derived macrophages. A Flow cytometry analysis and quantification of the expression of M2 markers CD206 and CD163, as well as M1 markers CD80 and CD86, in LPS-stimulated macrophages pretreated with E2 in the presence or absence of the SGK1 inhibitor GSK650394. The plot show one representative flow cytometric analysis. B Transcript levels of ARG1 and IRF4 (differentiated M2 markers) were assessed using qRT-PCR in LPS-trigged macrophages pretreated with E2 in the presence or absence of GSK650394. C Concentrations of IL4 and IL-5 (Th2 cytokines), and IFN-γ (Th1 cytokine) in the supernatants of LPS-stimulated macrophages pretreated with E2 in the presence or absence of GSK650394. D Quantification of the arithmetic mean (SEM) ratio of IL-4/IFN-γ (Th2/Th1 cytokines) obtained from data presented (C). qRT-PCR analysis showing the mRNA levels of MMP9 (E), and VEGFA (F) in the presence or absence of GSK650394 in LPS-stimulated macrophages pretreated with E2. GAPDH served as the internal control. Data are presented as the arithmetic means ± SEM for three individual experiments. *P < 0.05, **P < 0.01, ***P < 0.001, contrasted with the control group; ∆P < 0.05, ∆∆P < 0.01, ∆∆∆P < 0.001, contrasted with the LPS group; #P < 0.05, ##P < 0.01, ###P < 0.001, contrasted with the LPS + E2 group. SGK1, serum-glucocorticoid regulated kinase; E2, estradiol; ARG1, arginase 1; IRF4, immune regulatory factor 4; LPS, lipopolysaccharide; IL, interleukin; IFN-γ, interferon gamma; MMP9, matrix metalloproteinase 9; VEGF-A, vascular endothelial growth factor A; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; SEM, Standard Error of the Mean