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In brief, HUVECs were lysed in Co-IP buffer containing 20 mM Tris-HCl (pH 7.5), 100 mM NaCl, 0.5 mM EDTA, 0.5% NP-40, 0.5 mM PMSF, and 0.5% protease inhibitor cocktail on ice for 20 min

To monitor LC3-II puncta, we also subcloned Mytho in a PBI3xFlag vector that contains a YFP-LC3 expression gene in a different cloning site (PBI YFPLC3-3xFlagMYTHO, Addgene) using KOD Hot Start DNA polymerase

This is supported by observations that glucagon can stimulate insulin release 17,18,19,20 , and that cells also produce glucagon-like peptide-1 (GLP-1) 21,22,23,24,25

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Learn about the different ways of disposing LiPo batteries in this article: Never to puncture LiPo batteries, as doing so can cause a fire