Fig. 1. KMS99220 activates the AMPK signaling pathway in microglial cells. (A) AMPK activity in the presence of KMS99220 was measured using a radiometric assay as described in Methods. The data are expressed as % of untreated control±SEM; ^#p<0.05 and ^##p<0.01. (B) BV2 cells were treated with 1, 5 and 10 µM KMS99220 for 15 min. (C, D) BV2 cells were treated with 10 µM KMS99220 for 15, 30, 60 and 180 min. Western blot analysis was performed against p-AMPK, AMPK, β-actin and Lamin B. For quantitation, values obtained from densitometry were normalized against respective loading controls (AMPK for B and C; β-actin or Lamin B for D) and are expressed as induction fold of untreated control, as indicated above each panel. (D) Nuclear and cytoplasmic extracts were subjected to western blot analysis against p-AMPK. Lamin B was used as a loading control and nuclear marker. (E) BV-2 cells were exposed to various concentrations of KMS99220 for 24 h, and cytoxicity was assessed by ATP assay.