Hypothalamic AMPK regulates Npy and Pomc expression in response to low glucose availability. (A) Npy and Pomc mRNA expression levels in hypothalamic neuronal cells were measured by quantitative real-time PCR. N41 and N43/5 cells were treated with AICAR (0.5 mM) for 12 h (*, p< 0.05; **, p < 0.01 for vehicle [Veh] vs. AICAR; N41: n = 6, N43/5: n = 9). (B) Levels of Npy and Pomc mRNA in N41 and N43/5 cells treated with 2DG (20 mM) or glucose-free medium (0 mM glucose) with or without compound C (Com C; 16 μM) for 4 h (**, p < 0.01; ***, p < 0.001 for vehicle [Veh] vs. compound C; ##, p < 0.01; ###, p < 0.001 for 25 mM glucose vs. 2DG or glucose-free medium in vehicle-treated groups; n.s., not significant for 25 mM glucose vs. 2DG or glucose-free medium in compound C-treated groups; N41: n = 7 or 8, N43/5: n = 6-8). (C) Prkaa1 and Prkaa2 mRNA expression levels in hypothalamic neuronal cells were measured by quantitative real-time PCR. Cells were infected with lentiviruses containing control (shCont) or both Prkaa1 and Prkaa2 (shAmpk) shRNAs (###, p < 0.001 for shCont vs. shAmpk; N41: n = 6, N43/5: n = 8). (D) Npy and Pomc mRNA expression levels in hypothalamic neuronal cells measured by quantitative real-time PCR. Cells were infected as in (C) and then 2DG or glucose-free medium was given for 4 h (**, p < 0.01; ***, p < 0.001 for shCont vs. shAmpk; ##, p < 0.01; ###, p < 0.001 for 25 mM glucose vs. 2DG or glucose-free medium in the shCont group; ††, p < 0.01; †††, p < 0.001; n.s. for 25 mM glucose vs. 2DG or glucose-free medium in the shAmpk group; N41: n = 6, N43/5: n = 4–6).