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Figure 2


Fig. 2. Shear flow negatively regulates beta-catenin signaling and cell cycle. (A) SW480 cells transiently transfected with TOPFLASH reporter were exposed to shear stress (0-35 dyn/cm2) for 12 hours. Inset in A, western blot of beta-catenin in SW480 cell lysates before and after shear flow. (B) SW480 cells transiently transfected with cyclin D1 reporter gene (with or without TCF sites) were exposed to shear stress (0-35 dyn/cm2) for 12 hours. (C) SW480 cells plated at low (800,000 cells) and high (6 million cells) density were transiently transfected with TOPFLASH and exposed to shear stress of 15 dyn/cm2 for 12 hours. (D) SW480 cells transiently transfected with TOPFLASH were exposed to shear stress of 15 dyn/cm2 for 0 to 24 hours. (E) SW480 cells transiently transfected with TOPFLASH were exposed to shear stress (15 dyn/cm2) for 12 hours and incubated in static culture for 0 to 25 hours. (F) A1N4 cells, transiently transfected with TOPFLASH, FOPFLASH or pcDNA, and beta-catenin were exposed to shear stress of 15 dyn/cm2 for 12 hours. (G,H) Control cells (G) or cells exposed to shear flow for 24 hours (H) were analyzed by FACS. Experiments were performed in triplicate with consistent and repeatable results.





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