Figure 6.

Knockdown of AMPK in the ARC decreases autophagy, food intake and body weight in mice fed ND or HFD. (A) Schematic illustration of the surgical coordinates and expression of eGFP delivered by lentiviruses targeting the arcuate nucleus (ARC). Scale bar: 200 μm. Daily food consumption and body weight changes in male mice fed (B) a normal diet (ND) and (C) a high-fat diet (HFD) after lentivirus injection for the indicated days (*, p < 0.05; **, p < 0.01; ***, p < 0.001 for the control shRNA group [shCont] vs. AMPK shRNA [shAmpk] group; ND: n = 20; HFD: shCont group, n = 18; shAmpk group, n = 23). (D) Relative levels of total AMPK, p-ULK1 (Ser555), total ULk1, p-RPTOR (Ser792), total RPTOR, NBR1, LC3, and GAPDH in the ARC tissue from the shCont and shAmpk groups fed a HFD were examined by immunoblotting. p-ULK1 and p-RPTOR were normalized to corresponding total proteins. Other protein levels were normalized to GAPDH. Protein levels in the control shRNA group were arbitrarily set to 1.0 (*, p < 0.05; **, p < 0.01 for shCont vs. shAmpk; n = 5). (E) Npy and Pomc mRNA expression levels in the ARC from shCont and shAmpk groups fed a HFD measured by quantitative real-time PCR. (*p < 0.05 for shCont group vs. shAmpk group; n = 8).