Figure 5.
(A) In vitro actin polymerization assay. Compounds 3 and 4 (used at the concentration of 5 µM) were incubated with the labeled rabbit muscle actin in order to verify their ability to inhibit the protein polymerization. (B) In vitro actin depolymerization assay. After actin polymerization, compounds 3 and 4 (5 µM) were added to the reaction mixture, in order to determine their ability to act as depolymerizing agents. For both the assays, the vehicle DMSO was used as a negative control. Actin-targeting agents, LA and CB, both at the concentration of 5 μM, were used as positive controls. The assemblage of the actin filaments was established by monitoring the fluorescence (λEx/Em= 365/410 nm) in kinetic mode for 1 h at room temperature by using a microplate reader. The graphics are representative of three separate experiments and error bars represent the standard deviations.