Autophagy Analysis in Colon Cancer Cells

Stable colon cancer SW480 and DLD1 cell lines were generated expressing DsRed-LC3-GFP reporter.^{18 (link)} After cultured under different conditions, cells were fixed and fluorescent signals of DsRed and GFP were detected using an Olympus FlowView FV1000 confocal laser-scanning microscope (Waltham, MA, USA). The autophagy index was calculated as the relative fluorescence intensity of DsRed to that of GFP obtained from flow cytometry analysis.

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Wen Y.A., Xing X., Harris J.W., Zaytseva Y.Y., Mitov M.I., Napier D.L., Weiss H.L., Mark Evers B, & Gao T. (2017). Adipocytes activate mitochondrial fatty acid oxidation and autophagy to promote tumor growth in colon cancer. Cell Death & Disease, 8(2), e2593-.

Publication 2017

FlowView FV1000

Autophagy Cell lines Cells Colon cancer Confocal laser scanning microscope Cultured different Flow cytometry Fluorescence

Corresponding Organization : University of Kentucky

Other organizations : Markey Cancer Center

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Variable analysis

independent variables

Different conditions

dependent variables

Autophagy index (relative fluorescence intensity of DsRed to GFP)

control variables

Stable colon cancer SW480 and DLD1 cell lines expressing DsRed-LC3-GFP reporter
Cells were fixed
Fluorescent signals of DsRed and GFP were detected using an Olympus FlowView FV1000 confocal laser-scanning microscope

controls

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

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