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. 2020 Mar 6;10:48. doi: 10.3389/fcimb.2020.00048

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Copyright © 2020 Iraqi, Edri, Greenshpan, Kundu, Bolel, Cahana, Ottolenghi, Gazit, Lobel, Braiman and Porgador.

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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Site-specific mutations, disrupting the N-linked glycosylations in GP, do not affect co-localization of GP with HLA-A*0201. Cells were stably transfected with either CFP-fused HLA-A*0201 (A,C) or CFP-fused NKp46 (B,D) and then transiently transfected with either YFP-fused wild type GP (A,B) or YFP-fused double mutant GP (C,D). Thirty hours post transfection, cells were assayed by confocal microscopy using a 40 × /1.3 UPLSAPO objective (magnification including ocular lens is 400 ×). Summary plot shows FRET efficiency in various samples. Results are representative images from 50 images taken from the experiment. Scale bar (20 μm) is shown at the bottom right corner of each plot. (E) Positive control, (F) Negative control, (G) Summary results. Graphs represent mean ± SEM. ***p < 0.001, ordinary one-way ANOVA multiple comparisons test.