Figure 9.

Adropin^34–76 treatment suppresses glucose production in primary mouse hepatocyte. A, glucose production from the hepatocytes was determined by quantifying glucose levels in culture media. The assay was performed from three hepatocyte preparations, and the data were pooled and presented as a percentage of the vehicle-treated values (n = 10). The levels of glucose production in the vehicle-treated group were around 0.1 mg/mg of protein/h. B, cAMP levels in HEPG2 liver cells were measured in the presence of increasing levels of forskolin (an activator of adenylate cyclase) in the culture media. The experiments were repeated three times. C, the phosphorylation levels of Ser¹³³ in CREB and total CREB levels, and the phosphorylation levels of PKA substrates in the hepatocytes were determined by Western blotting (n = 2). Heat shock protein 90 (Hsp90) was used as the loading control. D, the message levels of glucose production genes, including G6Pase (G6pc) (n = 5) and PEPCK (Pck1) (n = 2–3), in the hepatocytes were determined by real-time PCR. The quantitation of Pck1 was repeated in another experiment (n = 3), and the levels of Pck1 in the adropin-treated group were below the detection limit. Hypoxanthine guanine phosphoribosyltransferase was used as the reference gene. *, p ≤ 0.05, adropin versus vehicle. Error bars, S.E.