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. 2006 Sep;18(9):2275–2293. doi: 10.1105/tpc.105.040279

Figure 7.

Figure 7.

SYP21 Overproduction Perturbs PVC Morphology and Inhibits Recycling of the Plant Vacuolar Sorting Receptor.

(A) Transient expression experiment (16-h incubation) to document normal appearance of individual Golgi bodies highlighted with the marker ST-GFP (in green) and PVC bodies highlighted by YFP-BP80 (in red). Shown is a cortex view of the protoplast. Images in bright field (DIC) are shown at the right. Note that ST-GFP–labeled Golgi bodies are frequently weakly labeled by YFP-BP80 (arrows). Bar = 5 μm.

(B) and (C) Overexpression of YFP-SYP21 (16-h incubation) does not influence the morphology of Golgi bodies in the cells while PVCs form large clusters. Note that ST-GFP–labeled Golgi bodies are not labeled by YFP-SYP21 and that there is no bleeding through of YFP signals in the GFP channel. Bars = 5 μm.

(D) Statistical analysis of the occurrence frequency of Golgi bodies (white bars) and PVC bodies (gray bars). The frequency is shown in percentage of the total number of punctate signals in a sample, and data are from 15 independent samples ± se in each case, scoring 762 individual punctate structures for YFP-BP80/ST-GFP and 805 for YFP-SYP21/ST-GFP. Note a clear shift to lower PVC numbers relative to Golgi numbers when YFP-SYP21 is used as PVC marker.

(E) Statistical analysis of the occurrence frequency for PVC bodies highlighted by either YFP-BP80 or YFP-SYP21 that are smaller than 1 μm (white bars), between 1 and 2 μm (gray bars), and above 2 μm (black bar). The frequency is shown as a percentage of the total number of punctate signals in a sample, and data are from nine independent samples ± se (386 punctate structures scored in total) for YFP-BP80 and 30 independent samples (262 punctate structures scored in total) for YFP-SYP21. Note a clear shift to large PVCs when YFP-SYP21 is overexpressed.

(F) and (G) Two examples of PVC clusters highlighted by YFP-SYP21, shown at high magnification. Notice that SYP21 overexpression causes the formation of clusters that appear to be composed of distinct PVC bodies tethered together. Bars = 5 μm.

(H) to (J) Still images of Supplemental Movie 1 online, showing the first, middle, and last frames. Note the variable shape of the PVC clusters highlighted by the arrows. Bar = 5 μm.

(K) Transient expression experiment showing subcellular localization of cotransfected YFP-SYP21 and GFP-BP80. Notice the almost complete colocalization of the two PVC markers and the enlarged structures (arrows). Bar = 5 μm.