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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.