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Additional Supporting Information may be found in the online version of this article.

FilenameFormatSizeDescription
CM_21057_sm_SuppFigS1.tif3810KSupporting 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.tif5181KSupporting 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.mov8823KSupporting 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.mov327KSupporting 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.mov10555KSupporting 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.mov11186KSupporting 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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