Metformin's inhibition on AGEs-induced NFκB signaling is AMPK dependent. BMDMs were divided into 4 groups: control, AGEs, AGEs + MET, and AGEs + MET + CC group. In AGEs group, cells were cultured with AGEs at 200 mg/L for 60 min; in AGEs + MET group, cells were pretreated with metformin for 60 min and then cultured with AGEs at 200 mg/L for 60 min; in AGEs + MET + C-C group, cells were pretreated with Compound C, an AMPK inhibitor, at 5 μM for 60 min, and then they were treated with metformin at 2.0 μM for 60 min followed by AGEs at 200 mg/L for 60 min; in control group, cells were cultured with BSA at 200 mg/L for 60 min. p65 nuclear translocation of each group was evaluated by immunofluorescent staining. Primary antibodies against p65 and Cy3 (red) labeled secondary antibodies were used to detect p65; DAPI (blue) was used to stain the nucleus. Bar graphs represent the results (mean ± SD) of three independent experiments. Bar = 50 μm. One-way ANOVA was applied and the overall ANOVA was significant. ∗∗
p < 0.01 and ∗∗∗
p < 0.001 when compared between selected groups.