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. 2016 Sep 28;2016:4847812. doi: 10.1155/2016/4847812

Figure 3.

Figure 3

Metformin activates AMPK and inhibits AGEs-induced RAGE expression and NFκB activation. (a) BMDMs were divided into 4 groups: control, AGEs, MET, and AGEs + MET group. In AGEs group, cells were cultured with AGEs at 200 mg/L for 24 h; in MET group, cells were cultured with metformin at 2.0 μM for 24 h; in AGEs + MET group, cells were pretreated with metformin for 60 min and then cultured with AGEs at 200 mg/L for 24 h; in control group, cells were cultured with BSA at 200 mg/L for 24 h. Western blot analysis was performed to measure protein levels of RAGE and phosphorylated AMPK (p-AMPK). Tubulin was used as internal control. (b) BMDMs were pretreated with or without metformin (2.0 μM) for 60 min before AGEs (200 mg/L) stimulation for different time intervals (0, 30, 60, and 180 min). Protein levels of NFκB-p65 (p65) and phosphorylated NFκB-p65 (p-p65) were measured by western blot. Tubulin was used as internal control. Bar graphs represent the results (mean ± SD) of three independent experiments. One-way ANOVA was applied and all the overall ANOVA was significant. # p > 0.05; ∗ p < 0.05; ∗∗ p < 0.01; and ∗∗∗ p < 0.001 when compared between selected groups.