Journal
MOLECULAR BIOLOGY OF THE CELL
Volume 27, Issue 5, Pages 848-861Publisher
AMER SOC CELL BIOLOGY
DOI: 10.1091/mbc.E15-09-0664
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Funding
- National Medical Research Council [NMRC/CBRG/007/2012]
- Ministry of Education [AcRF Tier1 RG 18/11, RG 48/13]
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Cellular functions of the Golgi are determined by the unique distribution of its resident proteins. Currently, electron microscopy is required for the localization of a Golgi protein at the sub-Golgi level. We developed a quantitative sub-Golgi localization method based on centers of fluorescence masses of nocodazole-induced Golgi ministacks under conventional optical microscopy. Our method is rapid, convenient, and quantitative, and it yields a practical localization resolution of similar to 30 nm. The method was validated by the previous electron microscopy data. We quantitatively studied the intra-Golgi trafficking of synchronized secretory membrane cargoes and directly demonstrated the cisternal progression of cargoes from the cis-to the trans-Golgi. Our data suggest that the constitutive efflux of secretory cargoes could be restricted at the Golgi stack, and the entry of the trans-Golgi network in secretory pathway could be signal dependent.
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