Fig. 4

Testosterone-mediated FASN suppression inhibited NF-κB activation. A MG6 cells were stimulated with lipopolysaccharide (LPS) for 45 min in the presence or absence of C75 at the indicated concentrations. p65 phosphorylation was analyzed using western blot. The graph on the right shows the phosphorylated/total protein ratio of p65. B MG6 cells were stimulated with LPS for 45 min in the presence or absence of 50 nM testosterone. p65 phosphorylation was analyzed using western blot. Representative blots and the phosphorylated/total protein ratio of p65 are shown. C MG6 cells were transfected with control miRNA and indicated miRNA mimics. After 48 h, cells were stimulated with LPS for 45 min, and p65 phosphorylation was analyzed using western blot. Representative blots and the phosphorylated/total protein ratio of p65 are shown. D MG6 cells were stimulated with LPS for 2 h in the presence or absence of 50 nM testosterone. Tnfa expression was analyzed using qPCR. Data are expressed as mean ± SD from three independent experiments. E Secretion of TNF-α after 24 h of LPS stimulation in the presence or absence of testosterone was measured using enzyme-linked immunosorbent assay. Data are expressed as mean ± SD from three independent experiments. Full membrane images are depicted in Supplementary Figures S2–S4. *p < 0.05 versus the corresponding value for the indicated comparison according to one-way analysis of variance followed by Tukey–Kramer’s HSD test