(A) 7,8-dihydroxyflavone chemical structure with every position in each ring labeled. (B) Chemical Structures of flavonoids from Indofine, Inc. (C) Immunoblotting analysis with neuronal lysates. Primary rat cortical neurons from E17 embryos (13 DIV) were treated with 500 nM various chemicals for 15 min. The neuronal cell lysates were collected and resolved on 10% SDS-PAGE. Immunoblotting was conducted with various antibodies. p-TrkB Y817 antibody was employed at 1:20,000–40,000 dilution. (D). Phospho-Akt 473 ELISA. The cell lysates (20 μg/sample) from the neurons treated with various indicated drugs were analyzed by p-Akt ELISA. The quantitative p-Akt activity in ELISA correlated with TrkB activity. Results were expressed as mean ± SEM (*P<0.05, compared with the vehicle control group, Student t-test, n=3).