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. 2016 Nov 21;215(4):499–513. doi: 10.1083/jcb.201608123

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© 2016 Thomas and Fromme

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 4. — Sec7 activity is required for TRAPPII recruitment and Ypt31/32 activation at the late Golgi/TGN. (A, left) Localization of Trs130-mNeonGreen relative to the late Golgi marker Sec7-mRFPmars in cells treated with DMSO (vehicle control) or BFA for 15 min. (right) Recruitment of TRAPPII to Golgi compartments was measured by quantifying the ratio of Trs130-mNeonGreen fluorescence to that of Sec7-mRFPmars in Sec7-mRFPmars puncta. Error bars represent 95% CIs for n = 55 (DMSO) or n = 21 (BFA-treated) compartments. (B, left) Localization of Trs130-mNeonGreen relative to Sec7-6xDsRed in cells treated with DMSO (vehicle control) or MNTC for 10 min. (right) Recruitment of TRAPPII to Golgi compartments was measured as in A. Error bars represent 95% CIs for n = 55 (DMSO) or n = 42 (MNTC-treated) compartments. (C, left) Localization of Trs130-mNeonGreen in a sec7-1 mutant after shifting to the restrictive temperature for 20 min. (right) Line trace quantification of cytosolic and Golgi localization of TRAPPII. Error bars represent 95% CIs for n = 7 (30°C) or n = 9 (37°C) cells. (D, left) Localization of GFP-Ypt31 relative to Sec7-mRFPmars in cells treated with DMSO or BFA for 15 min. (right) Recruitment of Ypt31 to Golgi compartments was measured as in A and B. Error bars represent 95% CIs for n = 58 (DMSO) or n = 37 (BFA-treated) compartments. Bars, 2 µm. White arrowheads denote colocalization of TRAPPII or Ypt31 with Sec7 at Golgi compartments. ***, P < 0.001.