Fig. 2.
Trafficking defects in mtv5 mutants. (A) CLV3:T7:CTPPBL relocalizes in mtv5. Electron micrographs show that the chimeric protein is trafficked to the vacuole in the Vac2 parental line but trafficked to the apoplasm in mtv5. Arrows designate the locations of gold particles. V, vacuole; CW, cell wall. (Scale bars, 200 nm.) (B) Col-0, Vac2, and mtv5 protoplasts. Protoplasts were incubated for 24 or 48 hours as indicated. Cells and media were separated, and proteins were analyzed in a dot-blot by using an α-T7 antibody. (C) Trafficking of a transiently expressed Arabidopsis aleurain-like protein (AtALP:GFP) is not affected by the mtv5 mutation. The 70-kDa protein is the AALP:GFP fusion, and the 27-kDa protein is GFP that was released from the 70-kDa chimeric protein upon correct delivery to the vacuole. The experiment was carried out as in B, except that protein extracts were resolved using SDS/PAGE and visualized using an α-GFP antibody.