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. 2009 Dec 22;285(8):5488–5496. doi: 10.1074/jbc.M109.058024

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© 2010 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 3. — Generation and characterization of Ran Tg mice. A, pAC-Ran construct for Ran Tg mice generation. The 4.9Kb ClaI/ClaI fragment was used for microinjection. B, Southern blot genotyping of Ran founder tail DNA. The 2.38-kb band specific to the Ran transgene is indicated. C, real-time RT-PCR of Ran mRNA from spleen and lymph node cells. Mean ± S.D. of ratios of Ran versus β-actin signals are shown. Samples are in triplicate. D, Ran overexpression in Tg T cells according to immunoblotting. Cytosolic and nuclear proteins were fractionated from Tg and WT T cells and were analyzed for Ran protein expression using immunoblotting (20 μg/lane). The cytosolic and nuclear protein purity was verified by α-tubulin and histone H3 levels.