Analyzing Autophagy Markers in Colon Cancer

Human colon cancer cells were harvested and detergent-solubilized cell lysates were obtained as described previously.^{32 (link), 33 (link), 34 (link), 35 (link)} Equal amounts of cell lysates were resolved by SDS-PAGE and subjected to western blot analysis. The phospho-ACC, phospho-AMPK, ACC, AMPK, Beclin 1, LC3 and E-cadherin antibodies were obtained from Cell Signaling (Danvers, MA, USA). The vimentin and p62 antibodies were from BD Biosciences (San Jose, CA, USA). The γ-tubulin antibody was from Sigma-Aldrich.

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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

phospho-ACC phospho-AMPK Beclin 1 E-cadherin γ-tubulin antibody

Antibodies Antibody Beclin 1 Cell Colon cancer Detergent E cadherin Human Sds page Tubulin Vimentin Western blot analysis

Corresponding Organization : University of Kentucky

Other organizations : Markey Cancer Center

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Phosphorylation of ACC and AMPK
Levels of Beclin 1, LC3, E-cadherin, vimentin, and p62

control variables

Equal amounts of cell lysates

positive controls

None explicitly mentioned

negative controls

None explicitly mentioned

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