Figure 2

ATP depletion and activation of AMP-dependent protein kinase (AMPK) caused by inhibition of mGluR5. (A) BV-2 cells were incubated with 100 μM MPEP for 1, 2, 3 and 4 hours, followed by determination of cellular ATP levels. (B) Cells were treated for 2 hours with 100 μM MPEP, 1 μM of the AMPK inhibitor compound C, 1 mM of the AMPK activator AICAR or combinations, followed by determination of cellular ATP content. Results represent mean ± SD of three independent experiments. Significance was tested by one-way ANOVA followed by Tukey’s test. *P <0.05, ***P <0.005. (C) Immunoblot analyses were performed on lysates of BV-2 cells that had been treated with MPEP or AICAR (positive control) for the indicated time period. (D) Densitometric analysis of phospho-AMPK band normalized against β-actin. (E) Immunoblot analyses were performed on lysates of BV-2 cells that had been pre-incubated with either AICAR (1 mM) or compound C (1 μM) prior to incubation with MPEP for 2 hours. (F) Densitometric analysis of phospho-AMPK band normalized against β-actin. MPEP, 2-methyl-6-(phenylethynyl)-pyridine.