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. 2022 Mar 21;298(5):101852. doi: 10.1016/j.jbc.2022.101852

Figure 2.

Figure 2

Effects of Lusianthridin, AMP, or A-769662 on activation and protection from dephosphorylation of various AMPK mutant complexes. A–C, activation of various AMPK mutant complexes by Lusianthridin, AMP, or A-769662 determined using the HTRF KinEASE assay kit and displayed as the fold increase in AMPK activity (±SEM, n = 3). D–F, increasing concentrations of Lusianthridin (D), AMP (E), or A-769662 (F) were incubated with phosphorylated (Thr172) α2β1γ1 WT or α2β1γ1 S108A (S108A) in the presence of protein phosphatase 2Cα (PP2Cα) for 10 min at 30 °C. Phosphorylation of AMPKα was determined by Western blot analysis using the anti-phospho-Thr172 antibody. Phosphorylation was quantified and is displayed above a representative Western blot image and is the means ± SEM of at least three independent experiments. AMPK, AMP-activated protein kinase; HTRF, Homogenous Time-Resolved Fluorescence.