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. 2017 Jul 13;7:5296. doi: 10.1038/s41598-017-05680-8

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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IFT88 is a component of ER-derived COPII coated vesicles. (A) Representative immunofluorescence staining images of endogenous IFT88 and SEC13 in the perinuclear region of NIH3T3 cells. The Manders Overlap Coefficient (MOC) between IFT88 and SEC13 is 0.72 ± 0.093. (B) Representative immunofluorescence confocal images showing colocalization of exogenously expressed GFP-IFT88 with endogenous Sec31A. MOC, 0.61 ± 0.03. (C and D) Representative confocal images showing colocalization of endogenous IFT88 with exogenously expressed ER marker DsRed-ER or exogenously expressed GFP-IFT88 with endo-Calnexin. Arrows mark the positions of the insets. MOC, 0.55 ± 0.11 in (C) and 0.68 ± 0.085 in (D), respectively. (E) Western analysis of IFT88 and SEC13 present in four differential centrifugation fractions in NIH3T3 cells. (F and G) Co-immunoprecipitation of exogenously expressed FLAG-IFT88 and GFP-Sec13 (F) or FLAG-IFT88 and CFP-Sec31A (G) in 293T cells. The cell lysates were precipitated with anti-FLAG M2 affinity gel and probed with anti-GFP or anti-Sec31A antibody for Western blot. The statistics analysis was computed based on data from at least 3 independent experiments.