Novel Small-Molecule AMP-Activated Protein Kinase Allosteric Activator with Beneficial Effects in db/db Mice
Figure 3
ZLN024 has no effect on the ADP/ATP ratio in L6 myotubes and requires an upstream kinase.
(A) ZLN024 did not affect the mitochondrial membrane potential after incubation for 3 hr in L6 myotubes; CCCP (10 µmol/l) was used as a positive control. (B) ZLN024 did not change the ADP/ATP ratio after incubation for 3 hr in L6 myotubes, CCCP (10 µmol/l) was used as a positive control (n = 3). (C) ZLN024 stimulates AMPK and ACC phosphorylation in HeLa cells in which only CaMKKβ acts as an AMPK upstream kinase. ZLN024 was incubated for 3 hr with A-769662 (20 µmol/l) as a positive control. The ratio of the phosphorylation level to the protein level of AMPK and ACC was determined (n = 2). (D) The time course of the stimulation of AMPK and ACC phosphorylation by ZLN024 (20 µmol/l). A-769662 (20 µmol/l) was used as a positive control. The ratio of the phosphorylation level to the protein level of AMPK and ACC was determined (n = 2). (E) AMPK and ACC phosphorylation by ZLN024 is blocked by the CaMKKβ inhibitor STO609. HeLa cells were incubated with ZLN024 (20 µmol/l) for 3 hr with or without prior treatment with STO-609 (10 µg/ml) for 30 min. #, P<0.1, *, P<0.05, **, P<0.01 compared with the untreated control.