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. 2020 Nov 19;8:607080. doi: 10.3389/fcell.2020.607080

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Copyright © 2020 Costa, Remigante, Civello, Bernardinelli, Szabó, Morabito, Marino, Sarikas, Patsch, Paulmichl, Janáky, Miseta, Nagy and Dossena.

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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The ICln Ser67Ala-induced IClswell is unresponsive to variations in O-glycosylation of cellular proteins. IClswell activation was monitored in HEK 293 Phoenix cells transfected for 24 h with ICln Ser67Ala and the transfection marker dsRed as separate proteins and (A) pre-incubated for 1 h with 100 μM PUGNAc plus 5 mM glucosamine or 0.1% DMSO plus 5 mM D-glucose as the control or (B) co-transfected with OGT or (C) OGA-L and the transfection marker EGFP. The current density-to-voltage relationships determined in an extracellular hypotonic solution (left panels) and the current density-to-time relationships (right panels) of IClswell are shown. (n) refers to the number of cells. DMSO, dimethyl sulfoxide; EGFP, enhanced green fluorescent protein. OGT, O-GlcNAc transferase; OGA, O-GlcNAcase.