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. 2022 Jan 24;12:748303. doi: 10.3389/fimmu.2021.748303

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Copyright © 2022 Francisco, Billod, Merino, Punzón, Gallego, Arranz, Martin-Santamaria and Fresno

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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O intermedium lipopolysaccharide (LPS) induces hTLR4/hTLR2 heterodimerization. (A) HEK293T cells were transiently transfected with the plasmids hTLR4-YFP (acceptor) and hTLR2-CFP (donor). After transfection, cells were left unstimulated (NS) or were stimulated with O. intermedium LPS (1 µg/ml) for 10 min. Fluorescence resonance energy transfer (FRET) between YFP and CFP was measured in in vivo cells by sensitized emission fluorescence. Corrected FRET images (FRET_eff) are displayed, and FRET efficiency is shown as a color-coded scale of values between 0% and 100%. (B) Quantification of FRET-positive structures per total cell number as well as the area and mean fluorescence intensity of each punctate structure. The results are shown as mean ± SEM of two independent experiments (p < 0.001 between treated and untreated cells in punctuate structures/cell and FRET efficiency).