Fig. 4.

PEPR1 secretion, but not its endocytosis, depends on ARF-GEF. (A–C) Subcellular localization of PEPR1-GFP after treatment for 60 min with DMSO (A), BFA (50 µM) (B), and CHX (50 µM) + BFA (50 µM) (C). (D) Experiments performed after pretreatment with CHX for 1 h. Subcellular localization of PEPR1-GFP after 40 and 60 min of pep1 elicitation (100 nM, 10-s pulse) in the presence or absence of BFA (50 µM). pepr1pepr2 seedlings complemented with the pRPS5A::PEPR1-GFP construct were stained with FM4-64 (4 µM, 5 min, three washouts). (E) Subcellular localization of TAMRA-pep1 in the presence or absence of BFA. Seedlings expressing VHA-a1-GFP were treated with TAMRA-pep1 for 10 s, washed, kept in the presence or absence of BFA (50 µM), and imaged after 40 and 60 min. (Insets) Details of BFA bodies at 2.5× magnification. In all experiments, 5-d-old seedlings were used. (Scale bars: 10 µm.)