Fig. 4

Glycolytic inhibitors suppress LPS-induced IKKβ activation by modulating AMPK/mTOR signaling in microglial cells. a BV-2 microglial cells were pretreated with 2-DG (600–1000 μM) for 30 min, followed by LPS (200 ng/mL) treatment for 15 min. The expression of p-IKKα/β, p-IκB-α, and IκB-α was analyzed by Western blotting (upper), and the relative protein levels were quantified by densitometric analysis (lower). b BV-2 microglial cells were transfected with si-Scramble or si-HK2. After 48 h, the cells were stimulated with LPS (200 ng/ml) for the indicated time (0, 5, 10, 20, 30, 60 min), and then, the expression of HK2, p-IκB-α, and IκB-α was determined by Western blotting (upper), and the relative protein levels were quantified by densitometric analysis (lower). c BV-2 microglial cells were pretreated with 2-DG (600–1000 μM) for 30 min, followed by LPS (200 ng/mL) treatment for the indicated time. After 1 h, the expression of p-AMPK and p-mTOR was determined by Western blotting (upper), and the relative protein levels were quantified by densitometric analysis (lower). d BV-2 microglial cells were pretreated with 2-DG (600–1000 μM) for 30 min, followed by LPS (200 ng/mL) treatment for 16 h. The ADP/ATP ratio was calculated as described in the methods and materials section. Data are presented as mean ± S.D. (n=3) and are representative of results obtained from three independent experiments. *p < 0.05, **p <0.01, compared to the LPS group