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. 2013 Jan 29;288(13):8815–8825. doi: 10.1074/jbc.M112.440503

FIGURE 3.

FIGURE 3.

VPS29 is dispensable for VPS35 membrane recruitment. A, immunoblot analysis on total, cytosolic, and microsomal proteins from wild-type (WT) and vps29-3 plantlets using anti-VPS35a, anti-H+-ATPase, and anti-cytosolic FBPase antibodies. H+-ATPase and FBPase were used as markers of membrane and cytosolic fractions, respectively. Molecular mass markers are indicated on the left in kDa. B, total proteins extracted from dry seeds of wild type (WT), vps26a vps26b, vps35a vps35c, and two transgenic vps35a vps35c independent lines (#1 and #2) stably expressing the VPS35a-GFP fusion protein under the pVPS35a promoter. Extracts were analyzed by SDS-PAGE followed by Coomassie Blue staining of the gel. Transgenic lines exhibit a WT-like pattern of seed storage proteins compared with the mutants. p12S: precursor of 12S-globulin. Molecular mass markers are indicated on the left in kDa. C, confocal microscopy images illustrating the localization of VPS35a-GFP in the roots of vps29-4+/− (left) and vps29–4−/− (right) plants. Arrows indicate punctate compartments, and asterisks point to abnormally enlarged/vacuolated compartments labeled with VPS35a-GFP. Scale bars, 10 μm.