Chromosomal Tagging of Exocyst Complex

Chromosomal tagging of SEC5, SEC8, EXO70 and EXO84 by Green Fluorescence Protein (GFP) was performed as previously described [38 (link)]. Cells were grown to log phase at 25 °C. The cells were then fixed for microscopy as described previously [39 (link)]. F-actin was stained with Alexa Fluor 488-conjugated phalloidin (Molecular Probes Corp.). Fluorescence microscopy was conducted with microscope CTR6000 (Leica) equipped with a Plan-Apochromat 100x, 1.40 NA oil immersion objective lens. Images were taken using LAS AF 1.5.1 acquisition software (Leica).

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Liu Y., Zhang T., Sun D, & Luo G. (2019). The Cdc42 effectors Gic1 and Gic2 regulate polarized post-Golgi secretion. Cell & Bioscience, 9, 33.

Publication 2019

LAS AF 1.5.1 acquisition software

Alexa fluor 488 Cells Chromosomal F actin Fluorescence microscopy Green fluorescence protein Immersion Lens Microscope Molecular probes Phalloidin

Corresponding Organization :

Other organizations : China Medical University

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Variable analysis

independent variables

Chromosomal tagging of SEC5, SEC8, EXO70 and EXO84 by Green Fluorescence Protein (GFP)

dependent variables

Fluorescence microscopy of cells

control variables

Cells were grown to log phase at 25 °C
Cells were fixed for microscopy as described previously [39]
F-actin was stained with Alexa Fluor 488-conjugated phalloidin (Molecular Probes Corp.)
Fluorescence microscopy was conducted with microscope CTR6000 (Leica) equipped with a Plan-Apochromat 100x, 1.40 NA oil immersion objective lens
Images were taken using LAS AF 1.5.1 acquisition software (Leica)

controls

No positive or negative controls were explicitly mentioned in the provided information.

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