Fig. 2
AtEH/Pan1 localize to the PM as well as to punctate structures that are likely to be autophagosomes. a Representative SEM image of pollen from a heterozygous ateh1/pan1-1 plant showing normal and shrivelled pollen in a 1:1 ratio. b–e In the Arabidopsis root (e, f) and hypocotyl cells (l, m), AtEH/Pan1-mRuby3 proteins, in their respective complemented mutant background, localize predominantly to the PM at dynamic endocytic foci. To some extent, discrete larger punctae can be observed. f, g An antibody raised against AtEH2/Pan1 recognizes both proteins in Arabidopsis seedlings (g) and recognizes GFP-AtEH/Pan1 fusions transiently expressed in N. benthamiana. h–j Immunolocalization using the anti-AtEH/Pan1 antibody confirms the PM localization in Arabidopsis root cells and also identifies punctate labelling in leaf epidermal and root hair cells (Scale bar: 10 μm). k–o Punctate structures were identified in cells of Arabidopsis hypocotyl and cotyledons stably transformed with GFP-AtEH1/Pan1 (k, l). The same samples were further analysed at the ultrastructural level, and numerous double membrane autophagosome-like structures were identified (m–o; Scale bar: 500 nm)
