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Figure 3


Fig. 3. Immunofluorescence images illustrate that overexpressed, myc-tagged AtPex11a, - b, -c, -d and -e sort directly to peroxisomes during a 2.5 hour post-bombardment period in dual immunolabeled Arabidopsis and tobacco BY-2 suspension cells. The five myc-tagged AtPex11 proteins were introduced individually (biolistic bombardments) into Arabidopsis (A-J) or BY-2 (K-T) cells. Following bombardments (2.5 hours), cells were fixed in formaldehyde, cell walls perforated/digested with pectolyase (and cellulase, Arabidopsis only), and membranes permeabilized in Triton X-100. Cells were then dual immunolabeled with anti-myc plus anti-Cy-2-conjugated antibodies (1:500; 1 hour each) and anti-catalase plus RhodamineX-conjugated antibodies (1:2000; 1 hour each) (labeled columns of cells). Each representative image depicts one transformed cell per panel (labels on left side). In all cases, the myc-AtPex11 protein is colocalized with peroxisomal catalase (arrowheads point to examples in A,F,K,P). Arrows point to peroxisomes in neighboring nontransformed cells. Bars, 10 µm.





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