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. 2015 Mar 22;253(1):137–153. doi: 10.1007/s00709-015-0798-1

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© The Author(s) 2015

Open Access This article is distributed under the terms of the Creative Commons Attribution License which permits any use, distribution, and reproduction in any medium, provided the original author(s) and the source are credited.

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Fig. 4 — The localization of HvVPS60a-mCh differs in aleurone and subaleurone and shows plasma membrane localization in aleurone. a Single scans of barley aleurone and subaleurone cells showing transient transformation of HvVPS24-mCh in TIP3-GFP lines. In both, aleurone and subaleurone, HvVPS24-mCh localizes cytosolic and at small agglomerations (open arrowheads). Note the close ups to visualize co-localization. b Confocal single scans of HvVPS60a-mCh in TIP3-GFP lines and of co-transformation of HvVPS60a-mCh with TIP3-GFP. In aleurone, HvVPS60a-mCh shows strong signal at the pm (arrow), weak in the cytosol in parallel to small agglomeration structures (open arrowheads). Note the altered localization of HvVPS60a-mCh in subaleurone cells. HvVPS60a-mCh signal could be detected close to heterogenous, spherical PSVs (double arrow) but reduced at the PM (arrow). Note the close ups to visualize co-localization. c, d Localization of HvVPS24-mCh and HvVPS60a-mCh in aleurone and subaleurone. Relative number of cells showing localization in cytosol at agglomerations and at the PM was scored. Total number of cells: HvVPS24-mCh in aleurone (biological replicates 1; n = 18) and in subaleurone (biological replicates 3; n = 13); HvVPS60a-mCh in aleurone (biological replicates 1; n = 14) and in subaleurone (biological replicates 3; n = 24). The difference in the localization of HvVPS60a-mCh at the PM in aleurone and subaleurone is statistically significant (P < 0.05). Plus ends of the standard deviation are indicated. e Single scan of an aleurone cell showing co-localization of the plasma membrane marker PMA-EGFP (PE) and HvVPS60a-mCh at the pm (arrow). Fluorescence profile: X-axis, distance [μm] from 0 to 6 μm; Y-axis, intensity from 0 to 100. f Single confocal scan of a subaleurone cell that shows co-localization of HvVPS60a-mCh and the vacuolar membrane signal of the TIP3-GFP line. Note the double arrows indicating co-localization of HvVPS60a-mCh with the PSV membrane. Fluorescence profile: X-axis, distance [μm] from 0 to 8 μm; Y-axis, intensity from 0 to 100. Confocal single scans and z-series were made 24 h after bombardment. Scale = 5 μm