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Knock down of EB1 using short hair-pin RNA interference. B16F10 cells were transfected with EB1 knock down (KD) or MM control plasmids and stained with anti-EB1 monoclonal antibodies.  Non-transfected (NT) cells serve as an additional control. GFP co-expression identifies cells transfected with EB1 KD or MM control plasmids (lower panel). Whole cell EB1 fluorescence intensities are plotted relative to NT cells.

EB1 KD.jpg



IQGAP and WAVE localization at retracting and protruding edges.  (A) B16F10 cells were plated on laminin for 30 minutes then phase images were acquired at the indicated times (0-122 seconds). Kymographs represent retracting (R) and protruding (P) cell edges. (B) Cells were immediately fixed following live cell imaging and stained for actin, IQGAP1 and WAVE2. (C) Fluorescence intensity of actin (red), IQGAP1 (blue) and WAVE2 (green) at retracting and protruding edges are plotted.

IF-live cell.jpg



Microtubule targeting precedes dissociation of adhesions sites in B16F1 cells. Cells were co-transfected with GFP-paxillin and monomeric RFP-beta-tubulin subunit (mRFP-tubulin), and plated onto laminin.  Time is given in minutes and seconds. Arrows indicate a microtubule end targeting an adhesion site marked by GFP-paxillin.

MT targeting.jpg



Polymerization is required for EB1 association to microtubule plus-ends. Left: In control samples EB1 binds to the plus-ends of microtubules (inset, arrows). Right: In the presence of a known inhibitor (nocodazole for 30 sec) of microtubule polymerization EB1 binding to microtubule plus-ends is inhibited.

EB1 on MTs.jpg



A moving mouse melanoma cell. The cell was fixed and stained for actin.  Note numerous filopodia along the cell front.

B16F1 cell actin.jpg



Actin and nuclei in endothelial cells with z-stack.  A monolayer of endothelial cells was stained for actin and DNA. The z-series of images was deconvoluted before assembly into stack.

z series endothelial cells.jpg



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