Slides were prepared from isolated pancreata and visualized as previously detailed (11 (link)). ImageJ software was used to calculate the mean intensity on phospho-AMPK Thr 172 in the β-cell area and in the acinar tissue surrounding the islets. Western blotting was performed as previously described (11 (link)) with 100–150 human or mouse islets. Anti–acetyl-CoA-carboxylase (ACC),anti-phospho-ACC, anti-AMPK, anti–phospho-AMPK Thr 172, anti-Raptor, and anti–phospho-Raptor were from Cell Signaling Technology (Danvers, MA, USA). Anti-glucagon was from Sigma-Aldrich (St. Louis, MO, USA).
Martinez-Sanchez A., Nguyen-Tu M.S., Cebola I., Yavari A., Marchetti P., Piemonti L., de Koning E., Shapiro A.M., Johnson P., Sakamoto K., Smith D.M., Leclerc I., Ashrafian H., Ferrer J, & Rutter G.A. (2018). MiR-184 expression is regulated by AMPK in pancreatic islets. The FASEB Journal, 32(5), 2587-2600.
Acetyl coa carboxylaseGlucagon Human Mouse Pancreata Raptor Tissue β cell
Corresponding Organization : Imperial College London
Other organizations :
University of Pisa, Istituti di Ricovero e Cura a Carattere Scientifico, Vita-Salute San Raffaele University, Hubrecht Institute for Developmental Biology and Stem Cell Research, Leiden University Medical Center, University of Alberta, University of Oxford, Nestlé (Switzerland), AstraZeneca (United Kingdom)
Mean intensity on phospho-AMPK Thr 172 in the β-cell area
Mean intensity on phospho-AMPK Thr 172 in the acinar tissue surrounding the islets
Protein levels of acetyl-CoA-carboxylase (ACC), phospho-ACC, AMPK, phospho-AMPK Thr 172, Raptor, and phospho-Raptor
control variables
Previously detailed visualization method (11)
Previously described Western blotting protocol (11)
positive controls
Anti-glucagon antibody
negative controls
Not explicitly mentioned
Annotations
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