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CdGAP regulates cell migration and adhesion dynamics in two-and three-dimensional matrix environments†
Article first published online: 20 AUG 2012
Copyright © 2012 Wiley Periodicals, Inc.
Volume 69, Issue 9, pages 644–658, September 2012
How to Cite
Wormer, D., Deakin, N. O. and Turner, C. E. (2012), CdGAP regulates cell migration and adhesion dynamics in two-and three-dimensional matrix environments. Cytoskeleton, 69: 644–658. doi: 10.1002/cm.21057
- Issue published online: 18 SEP 2012
- Article first published online: 20 AUG 2012
- Manuscript Accepted: 19 JUL 2012
- Manuscript Revised: 18 JUN 2012
- Manuscript Received: 17 APR 2012
- NIH. Grant Numbers: RO1HL070244, RO1GM047607
Additional Supporting Information may be found in the online version of this article.
|CM_21057_sm_SuppFigS1.tif||3810K||Supporting Information Figure 1. Blebbistatin treatment induces the formation of small adhesions and a crescent morphology in GFP-cdGAP expressing cells. (A) U2OS cells transfected with GFP-cdGAP were spread on 10?g/ml collagen coated coverslips, treated with either DMSO vehicle control or blebbistatin, and stained for paxillin and actin. Insets demonstrate that GFP-cdGAP localizes with paxillin to both large adhesions in vehicle treated cells and small adhesions in blebbistatin treated cells.|
|CM_21057_sm_SuppFigS2.tif||5181K||Supporting Information Figure 2. CdGAP localizes to 3D matrix adhesions in U2OS cells. (A) U2OS cells were seeded into 3D CDMs and then fixed and stained for endogenous cdGAP, actin, and fibronectin. (B) U2OS cells were co-transfected with GFP-cdGAP and mRFP-paxillin, seeded into 3D CDMs, and then fixed and stained for fibronectin.|
|CM_21057_sm_SuppMovieS1.mov||8823K||Supporting Information Movie 1. U2OS cells treated with control or cdGAP siRNA randomly migrating on collagen. Phase contrast images acquired every ten minutes, with movies playing back at ten frames per second. CdGAP siRNA treated cells migrate at a high velocity and rapidly extend and retract lamellipodia.|
|CM_21057_sm_SuppMovieS2.mov||327K||Supporting Information Movie 2. U2OS cells over expressing GFP or GFP-cdGAP migrating on collagen. Transfected cells are marked with an asterisk (*). Phase contrast images acquired every ten minutes, with movies playing back at ten frames per second. The migration of cdGAP over expressing cells is inhibited and they slowly extend small lamellipodia.|
|CM_21057_sm_SuppMovieS3.mov||10555K||Supporting Information Movie 3. U2OS cells stably expressing zyxin-GFP treated with control or cdGAP siRNA migrating on collagen. Fluorescence images acquired every two minutes, with movies playing back at ten frames per second. CdGAP siRNA-silenced cells rapidly extend lamellipodia and quickly form and disassemble adhesions as they migrate.|
|CM_21057_sm_SuppMovieS4.mov||11186K||Supporting Information Movie 4. U2OS cells co-transfected with zyxin-GFP and either control vector or myc-cdGAP migrating on collagen. Fluorescence images acquired every two minutes, with movies playing back at ten frames per second. CdGAP over expressing cells slowly extend lamellipodia and have a large percentage of adhesions that remain stable for the duration of the movie.|
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