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. 2011 Jul 15;22(14):2423–2435. doi: 10.1091/mbc.E11-04-0306

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© 2011 Wendt et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 7: — E-cad regulates β1 integrin expression during 3D outgrowth. (A) D2.OR and D2.A1 cells were propagated in 3D cultures for 72 h in the absence (0) or presence of TGF-β1 (5 ng/ml), and subsequently were analyzed for the expression of the indicated proteins by immunoblotting. Data are representative of at least three experiments. Int, integrin; Cat, catenin. (B) Control (GFP), WT E-cad (E-cad), or ΔE-Ecad (ΔE)-expressing D2.A1 cells were grown in 3D culture before visualizing the expression of β1 integrin (β1 Int), E-cad, and actin by immunoblotting. Data are representative of at least three experiments. (C) D2.OR cells expressing either a scrambled (sc) or an E-cadspecific (shE) shRNA were cultured in 2D or 3D conditions, and subsequently were analyzed for the expression of β1 integrin (β1 Int), E-cad, and actin as a loading control. (D) D2.OR and D2.A1 cells expressing either a scrambled (sc) or two distinct β1 integrin shRNAs (shβ1#1 and #2) were analyzed by immunoblotting to visualize the expression of β1 integrin (β1 Int), E-cad, and actin as a loading control. (E) D2.A1 cells expressing E-cad (E-cad) or depleted for β1 integrin (shβ1) were grown under compliant 3D cultures and imaged using phase-contrast microscopy (100×). Afterward, the percentage of branching structures was quantified. Data are the mean (±SE) of nine random fields of view over three independent experiments. ND, none detected.