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. 2016 Aug 31;6:32473. doi: 10.1038/srep32473

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Copyright © 2016, The Author(s)

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(a) The functional unit of the Golgi apparatus is the mini-stack, a pile of flat membrane cisternae. At its entry side (the cis-side) the mini-stack receives secretory cargo from the endoplasmic reticulum and at its exit side (trans-side) this cargo is delivered to the trans-Golgi network. At the trans-Golgi network, cargos are packaged into transport carriers and sorted to their final destination within the cell. We have found that WPB size is determined by the structural status of the Golgi apparatus²¹. Mini-stacks, whose diameter ranges between 500 and 1000 nm, impart a limit to the size of the transiting vWF cargo, generating what we have defined vWF “quanta”. When the mini-stack are linked to form the structure known as Golgi ribbon (route 1), both short and long WPBs are formed, since vWF quanta can be co-packaged together at the trans-Golgi network. When the mini-stacks are unlinked (route 2), co-packaging of quanta is prevented and short WPBs are produced. The micrographs exemplify these conditions. HUVECs were transfected with vWF-GFP to label the WPBs being made during the treatment. In untreated cells (control cells) mini-stacks are linked into a ribbon (GM130) and WPBs of various sizes are generated. In cells with unlinked mini-stacks (route 2, obtained by treatment with nocodazole), only short (or mini-) WPBs are produced. Scale: 5 um. (b) WPBs have a typical cylindrical morphology and in untreated cultured HUVECs (as the Control in panel a) their length ranges between 0.5 and 5 μm²¹. The total area covered by WPBs indirectly measures the vWF they store. Morphological analysis indicates that long WPBs (defined as those longer than 2 μm; red-coded in the cartoon) represent a minor fraction of total number of organelles but account for a large fraction of the area they cover (bar graph, black vs red bar). Therefore, while a minority, long WPBs store a significant fraction of vWF. The quantifications displayed were obtained by High-throughput morphometric analysis (see text) of 4 sets (means ± SD) of untreated cells. Number of WPBs measured ~10⁵.