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. 2011 May 15;22(10):1677–1685. doi: 10.1091/mbc.E10-08-0677

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© 2011 Capaldo 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 6: — ZO-3 is required for cyclin D1 TJ localization and cyclin D1 protein stability. (A) SKCO-15 cells treated with siRNA directed against ZO-2 (top panel) or ZO-3 (bottom panel) and immunostained for Thr-286 cyclin D1 (green) and ZO proteins (red). Arrows highlight mitotic cells. (B) Sucrose density gradient of colonic epithelial cells after ZO-3 depletion. Cyclin D1–ZO-3 cosedimentation in TJ fractions is diminished after ZO-3 depletion. (C) Subcellular fractionation reveals differential cyclin D1 protein stability after Chx treatment (50 μM, 1 h). (D) Cyclin D1 shows lower protein stability in ZO-3 siRNA-treated cultures. SKCO-15 cell lysates were harvested at the indicated times up to 4 h posttreatment with Chx (50 μM). (B) Densitometric analysis of Western blot data shows cyclin D1 protein levels in SKCO-15 cells after Chx treatment (n = 3).