Myosin II regulates cell surface area and endocytosis in Oli-neu cells. (A+B) Western blot analysis of myosin (myo) IIA and IIB after siRNA knock down in Oli-neu cells as compared to actin as a loading control. (C) Control siRNA, siRNA directed against myosin (myo) IIA, myosin (myo) IIB or both were nucleofected into Oli-neu cells. The cells were subsequently cultured on CSPG coated coverslips and changes in cell surface area were quantified after 16 h (means ± SEM; n > 100 cells, n.s. not significant, ***p < 0,001). (D) Quantification of dextran uptake (30 min) in Oli-neu cells cultured on CSPG coated coverslips (means ± SEM; n > 100 cells, **p < 0,01, ***p < 0,001). (E) Quantification of dextran endocytosis in Oli-neu cells cultured on CSPG coated or control coverslips and treated with 10 μM Y27632 or 50 μM blebbistatin for 1 h (means ± SEM; n > 80 cells, *p < 0,05,***p < 0,001). (F) Quantitative analysis of the endocytosis of dextran in Oli-neu cells nucleofected with control siRNA or siRNA against both myosin IIA and IIB (means ± SEM; n > 80 cells from 2 independent experiments, ***p < 0,001). Scale bar 20 μm.
Kippert et al. BMC Cell Biology 2009 10:71 doi:10.1186/1471-2121-10-71