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. 2012 May 31;287(30):25370–25380. doi: 10.1074/jbc.M112.385658

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© 2012 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 2. — Overexpressed and endogenous Jade-1 interacts with NPHP4. A, 293T cells were transiently transfected with V5-tagged Jade-1 (V5.Jade-1) and either FLAG-tagged NPHP4 (F.NPHP4) or a control protein (F.EPS^1–225). Immunoprecipitation was performed using an anti-FLAG antibody. V5.Jade-1 was coimmunoprecipitated by F.NPHP4 but not F.EPS^1–225. Note that in the cytosolic IP lysates, Jade-1 levels were decreased by NPHP4, whereas in whole cell lysates taken from the same experimental culture dish prior to the IP procedure, Jade-1 levels were increased by NPHP4. Endogenous tubulin staining from whole cell lysates shows equal amounts. IB, immunoblot. B, in the reverse experiment, 293T cells were transiently transfected with V5.NPHP4 and either F.Jade-1 or F.EPS^1–225. Immunoprecipitation was performed using an anti-FLAG antibody. F.Jade-1 but not F.EPS^1–225 coimmunoprecipitated V5.NPHP4. C, 293T cells were transiently transfected with F.NPHP4 or a control protein. Immunoprecipitation was performed using an anti-FLAG antibody. Endogenous Jade-1 was detected in the precipitates of F.NPHP4 but not in the control lane. The expression of Jade-1 was found to be equal in the lysates. HC, heavy chain. Blots are representative from three experiments.