Autophagy Visualization and Quantification

Images were collected with a Nikon ES80 epi-fluorescence microscope
equipped with a CoolSnap CCD camera using NIS-Elements software with
constant exposure settings and processed equivalently in PhotoShop.
ATG8-GFP localization as a reporter for autophagy was as described
by Eisenberg et al.^{29 (link)} The percentage of
cells showing autophagy was based on counting at least 100 cells per
technical replicate, and the experiment was performed on two biological
replicates. Propidium iodide staining was performed as previously
described.^{29 (link)}

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Koselny K., Green J., Favazzo L., Glazier V.E., DiDone L., Ransford S, & Krysan D.J. (2016). Antitumor/Antifungal Celecoxib Derivative AR-12 is a Non-Nucleoside Inhibitor of the ANL-Family Adenylating Enzyme Acetyl CoA Synthetase. ACS Infectious Diseases, 2(4), 268-280.

Publication 2016

CoolSnap CCD camera

Autophagy Fluorescence Propidium iodide Replicate

Corresponding Organization :

Other organizations : University of Rochester

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Percentage of cells showing autophagy
Propidium iodide staining

control variables

Exposure settings (constant)
Image processing (equivalently in PhotoShop)

controls

Positive control: ATG8-GFP localization as a reporter for autophagy, as described by Eisenberg et al.
Negative control: Not specified

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