Fig. 2

Myricetin inhibits the demethylation activity of KDM4A, KDM4B, and KDM4C. A–C Kinetic analyses of myricetin that inhibited the demethylation activity of KDM4A (A), KDM4B (B), and KDM4C C with increasing concentrations of α-KG (kinetics, the first raw; and the corresponding Lineweaver–Burk plot, the second raw), or with increasing concentrations of the H3K9me3 peptide (kinetics, the third raw; and the corresponding Lineweaver–Burk plot, the fourth raw). D Inhibition of H3 demethylation by KDM4A, KDM4B, or KDM4C in the presence of 50 μM myricetin using Western blotting analysis. The reaction mixture containing 10 μM enzyme, 100 μM inhibitor or blank buffer, and 5 μM of H3 in 50 mM HEPES, pH 7.5, 1 mM αKG, 2 mM ascorbate, and 50 μM Fe(II) was incubated at 37 °C for 30 min, followed by western blot analysis. H3 lysine modifications were probed with H3K9me3 and H3K36me3 antisera, respectively. M, myricetin