Inhibition of myosin light chain phosphorylation in MDA-MB-231 breast cancer cells by BDP5290. (A) MDA-MB-231 human breast cancer cells were treated with indicated concentrations of BDP5290 or Y27632 for 30 minutes prior to cell lysis and quantitative western blotting with indicated antibodies. (B) Inhibition of MLC phosphorylation by BDP5290 and Y27632. All results are shown as mean ± standard error of n = 3 independent replicates. (C) Immunofluorescence imaging of MDA-MB-231 cells that were untreated or treated with 0.5 μM Y27632 or BDP5290 for 30 minutes. After fixation and permeabilization, cells were stained with phalloidin to visualize filamentous actin structures and pMLC (S19) primary antibody as indicated.