Fig. 2.
The rank potency of flavonoid derivative–induced HEK293 cell detachment correlates with electron transport chain complex I inhibition. (A) Adherent HEK293 cells were grown for 24 hours in the presence of the indicated concentrations of the five flavonoids (DHM, mundulone, rotenone, deguelin, isorotenone). Detached cells were removed and the relative levels of adhered cells remaining were measured by proxy assay of PicoGreen quantification of total DNA. (B) Isolated mitochondria were supplied with NADH and an electron acceptor (Q1), and the loss of NADH over 5 minutes at 340 nM was measured in the presence of the indicated amounts of each flavonoid. IC50 values were calculated from monoexponential association curves fitted using GraphPad Prism. R.F.U., relative fluorescent units. Error bars are the average ± the s.d. of three experimental replicates.