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. 2016 Oct 15;27(20):3143–3155. doi: 10.1091/mbc.E16-05-0272

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© 2016 Rodrigues et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 8: — Changes to Arf1 localization and Golgi structure over the cell cycle. (A) Single sections of the subapical and nuclear regions of fixed Arf1-GFP–expressing embryos at interphase, metaphase, and telophase. At interphase, Arf1-GFP localizes to Golgi puncta and at the PM to a lesser extent and is excluded from the nucleus. At metaphase, Arf1-GFP increases at the PM (blue arrows) and ER (yellow arrows; inset, RFP-KDEL as a coexpressed ER marker) but is excluded from the spindle region within the ER. By telophase, the Arf1-GFP distribution resembles that at interphase. Side images are single sections below the nuclei of Arf1-GFP embryos at interphase, metaphase, and telophase stained for LqfR to show Golgi ministacks throughout the cell cycle. (B) 3D reconstructions of Arf1-GFP puncta from time-lapse imaging of a single embryo at different stages of the cell cycle. Arf1-GFP puncta are color coded according to volume. (C) Histograms showing Arf1-GFP (left) and GM130 (right) puncta volume distributions at different stages of the cell cycle (color coded). From interphase to telophase, puncta volumes decrease for both Arf1-GFP and GM130. The Arf1-GFP histogram is for time-lapse imaging of a single embryo, and each dot represents the percentage of Arf1-GFP puncta within a volume range (the same trend was observed for three embryos; unpublished data). The GM130 histogram is from imaging of fixed embryos at different cell cycle stages, and each dot represents a mean ± SD of the percentage of GM130 puncta within a volume range for the embryo at that cell cycle stage (embryo numbers in inset). Insets quantify total numbers of puncta per cell cycle stage (color coded). For Arf1-GFP, each bar represents one time frame (30 s/time frame) from time-lapse imaging of one embryo (the same trend was observed for three embryos), and for GM130, the bars show means ± SD. An increase in total puncta number was evident for both Arf1-GFP and GM130 (*p < 0.05). N, embryo numbers (compiled from two or more experiments, each with similar results). (A–C) Constructs expressed as in Figure 1.