Panels a-c, equivalent amounts (50μg) lysates harvested from LMVEC transfected with non-silencing siRNA or p190-directed siRNA were resolved by SDS-PAGE and immunoblotted for indicated protein. Membranes were stripped and reprobed for actin. Equivalent numbers of LMVEC were transfected with fluorescein conjugated non-silencing siRNA (panels d-g), Alexa-546 conjugated p190 siRNA (panels h-k), or Alexa-546 conjugated p190 siRNA plus p190-B siRNA (panels l-o) and plated onto collagen coated glass cover slips. At 96h posttransfection, the endothelial cells were exposed to vehicle or 5μM rottlerin for 30min. The endothelial cells were fixed and immunoflurorescently stained for filamentous actin using phalloidin (panels e, g, i, k, m, o). Nuclei were counter-stained with DAPI. Endothelial cells were visualized via fluorescence microscopy. Arrowheads indicate endothelial cells which took up fluorescently tagged siRNA. Experiments were performed in duplicate, and 4−5 images were obtained on each slide. Representative images are shown. Scale bars = 20μm. n = 3.