Fig. 4

AA suppresses METH-stimulated TNFα and IL-6 production through inhibition of NF-κB/STAT3 and MAPK-ERK signaling pathways in microglia BV2 cells. BV2 cells were pretreated with 20 μM AA or various inhibitors for 1 h and then stimulated with 1 mM METH for 24 h. a 20 μM AA suppresses 1 mM METH-induced translocation of NF-κB/STAT3 and phosphorylation of JAK2/ERK in BV2 cells. β-actin and lamin B were used to confirm equal sample loading. b Inhibitors of NF-κB (20 μM Bay11-7085), STAT3 (20 μM S3I-201), ERK (20 μM PD98059), or 20 μM AA strongly suppressed METH-induced TNFα and IL-6 production in extracellular levels. DNA-binding activity of NF-κB (c) and STAT3 (d) in nuclear extracts was measured by EMSA. Immunoblotting and EMSA were quantified by densitometric analysis. The data are representative of three independent experiments and quantified as mean values ± SEM (n = 4 to 6). Tukey’s multiple comparison test, *p < 0.05 compared to normal control, ^† p < 0.05 compared to METH treatment