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. 2007 May 15;104(21):9012–9017. doi: 10.1073/pnas.0703033104

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© 2007 by The National Academy of Sciences of the USA

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Fig. 5. — The Y442F mutant reduces Rho-GTP but is deficient biologically. (A) Rho-GTP in H358 cells and stable clones expressing DLC1, Y442F, or R718A were analyzed by Rhotekin pull-down assay followed by anti-RhoA blotting (Top). The expression of DLC1 in the stable clones and total RhoA loading controls were confirmed by immunoblotting. These H358-derived cells were used for the biological experiments in the figure. (B) Transwell migration assay. After 3 days, migrated cells that had come through the transwell filter were photographed after staining and then solubilized and quantified colorimetrically. The data are the mean ± SD of triplicate well measurements from one representative experiment. (C) Anchorage-independent growth assay. Cells were grown in soft agar (0.4%) and photographed after 4 weeks. The quantitative data are the mean number of colonies (± SD) >400 μm in diameter from one representative experiment.