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. 2019 Jan 18;9:792. doi: 10.3389/fendo.2018.00792

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Copyright © 2019 Asadi and Dhanvantari.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Glucagon and GRP78 directly interact and are localized within secretory granules in α-TC1-6 cells. (A) Western blot showing GRP78 immunoreactivity in: total cell extracts from untransfected (lane 2) and transfected (lane 3) cells; affinity-purified Fc-glucagon from isolated secretory granules (lane 4); and affinity-purified Fc alone from isolated secretory granules (lane 5). GRP78 binds to Fc-glucagon, but not Fc alone. (B) Immunofluorescence microscopy of glucagon (green), GRP78 (red) and both images merged. Cells were cultured on collagen-coated coverslips for 24 h in DMEM containing 25 mM glucose. Images were acquired, 2D deconvoluted and analyzed with NIS-Elements, software (Nikon, Canada). Pearson correlation coefficient (PCC) indicates strong correlation between GRP78 and glucagon (PCC = 0.85 ± 0.08). ROI shows areas of colocalization of GRP78 and glucagon within secretory granules.