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. 2013 Oct 11;25(10):4061–4074. doi: 10.1105/tpc.113.116012

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© 2013 American Society of Plant Biologists. All rights reserved.

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Figure 6. — NLA-Mediated Ubiquitination Triggers Clathrin-Dependent Endocytosis of PHT1;1 and PHT1;4 from the PM and Sorting into the Endosomal Trafficking Pathway.

(A) The punctate signals of YFP (arrowheads) were observed when PHT1;1-YFP or PHT1;4-YFP was coexpressed with NLA but not with NLA^C279A in tobacco leaves. Bars = 10 μm.

(B) The punctate structure of PHT1;1-YFP caused by NLA was diminished by applying Tyr A23, an inhibitor of clathrin-dependent endocytosis, but not by its analog, Tyr A51. The signal was observed 24 h after the chemicals were applied. Bars = 10 μm.

(C) and (D) The effect of E-64d on the localization of PHT1;1-YFP in the wild type (C) and nla-2 mutant plants (D). The samples were observed 3 h after treatment with E-64d and FM4-64. DMSO was used as the negative control. The aggregation of PHT1;1-YFP in the wild type (C) is indicated by arrowheads. Bars = 20 μm.