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. 2013 Oct 3;9(10):e1003659. doi: 10.1371/journal.ppat.1003659

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© 2013 Du et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) Confocal image of the subcellular localization of NbMIP1.1a in leaf epidermal cells. YFP-NbMIP1.1a was transiently expressed in leaves of N. benthamiana via agroinfiltration and imaged at 48 hpi using a Zeiss LSM 710 laser scanning microscope. YFP signal revealed that NbMIP1.1a is present in the cell membrane, cytoplasm and nucleus. PCD3-1002: a CFP-tagged plasma membrane marker [81]. DAPI: staining for nuclei. Scale bar represents 20 µm. (B) YFP-NbMIP1.1a was found in both the soluble fraction and the membrane fraction (upper panel). Protein extracts were centrifuged at 100,000×g to produce crude soluble (S100) and microsomal (P100) fractions. Fractions were analyzed by western blot following separation by SDS-PAGE. The gels were probed using anti-GFP, anti-V-H-ATPase (vacuolar H-ATPase subunit, a vacuolar membrane marker) and anti-PEPC (phosphoenolpyruvate carboxylase, a cytosolic marker) antibodies as indicated.