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. 2014 Mar 3;204(5):793–806. doi: 10.1083/jcb.201309084

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© 2014 Chi et al.

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Figure 6. — Loss of Vps1 dynamin family GTPase impacts endosome number and morphology. (A) The indicated GFP-tagged endosomal proteins and Vps10 were visualized in wild-type and vps1Δ cells. Maximum projections of deconvolved z-stacks are shown. Bar, 5 µm. (B) Quantitation of SNX-BAR–, retromer-, and Sec7-decorated (Golgi) compartments in wild-type and vps1Δ cells. The number of puncta per cell was determined by masking each new punctum in each z-section. The average number of puncta per cell, wild-type vs. vps1Δ, and standard deviation, is shown for each of the indicated proteins. Every endosome marker showed significantly more puncta per cell: Vps17, 8.2 ± 2.4 vs. 14.8 ± 6.8; Mvp1, 7 ± 2.5 vs 11.1 ± 1.7; Vps26, 12 ± 2.5 vs. 21 ± 2.5; Sec7 (Golgi), 10.1 ± 1.4 vs. 10.9 ± 1.9; n = 30 for each strain, P < 0.0001 (unpaired t test), wild-type and vps1Δ, respectively. (C) To better visualize vacuole localization of Vps10 in vps1Δ cells, a single plane of the z-section containing vacuoles from the maximum projection shown in A is shown. Bar, 5 µm. (D and E) Thin-section electron micrographs of multivesicular endosomes in wild-type and vps1Δ cells (F and G) with endosomes highlighted in reference images at low magnification (D and F; bar, 500 nm) and shown at high magnification (E and G; bar, 100 nm). (H) Multivesicular bodies in vps1Δ cells are larger than in wild-type cells (unpaired t test, P = 0.0084; vps1Δ 193.5 nm ± 7.267, n = 28; wild-type 158.3 nm ± 9.179, n = 12).