Localization of Lipid Droplet Proteins

For localization of different LD proteins, strains carrying C-terminal GFP fusions to the protein examined were mated to strain AN117-4B carrying pRS426-mTagBFP-Spo20^51-91 as a prospore membrane marker (Neiman et al., 2000 (link); Huh et al., 2003 (link); Lin et al., 2013 (link)). The resulting diploids were sporulated and stained with 5 mM BODIPY-TR methyl ester (Life Technologies, Carlsbad, CA) to visualize LDs. Images were collected on a Zeiss AxioObserver Z.1 microscope. All images were processed using AxioVision 4.7 software.

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Ren J., Pei-Chen Lin C., Pathak M.C., Temple B.R., Nile A.H., Mousley C.J., Duncan M.C., Eckert D.M., Leiker T.J., Ivanova P.T., Myers D.S., Murphy R.C., Brown H.A., Verdaasdonk J., Bloom K.S., Ortlund E.A., Neiman A.M, & Bankaitis V.A. (2014). A phosphatidylinositol transfer protein integrates phosphoinositide signaling with lipid droplet metabolism to regulate a developmental program of nutrient stress–induced membrane biogenesis. Molecular Biology of the Cell, 25(5), 712-727.

Publication 2014

BODIPY-TR methyl ester AxioObserver Z.1 microscope

Bodipy Diploids Ester Membrane Microscope Protein c Proteins Strain

Corresponding Organization :

Other organizations : University of North Carolina at Chapel Hill, Texas A&M University, Stony Brook University, Emory University, University of Utah, University of Colorado Health, Vanderbilt University

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Variable analysis

independent variables

Strains carrying C-terminal GFP fusions to the protein examined

dependent variables

Localization of different LD proteins

control variables

Strain AN117-4B carrying pRS426-mTagBFP-Spo20^51-91 as a prospore membrane marker

controls

Positive control: Strain AN117-4B carrying pRS426-mTagBFP-Spo20^51-91 as a prospore membrane marker
Negative control: Not explicitly mentioned

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