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. 2018 Jun 28;9:1144. doi: 10.3389/fimmu.2018.01144

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Copyright © 2018 Lillico, Pemberton and Stafford.

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 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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Live-cell imaging of IpLITR 1.1b-mediated target interactions at different incubation temperatures. Rat basophilic leukemia-2H3 cells (3 × 10⁵) stably co-expressing IpLITR 1.1b and LifeAct-GFP were incubated at 37°C (A,B) or at 27°C (C–E) with 9 × 10⁵ αHA monoclonal antibody-coated 4.5 µm bright blue microspheres. Immediately after the addition of target beads, images were collected at 10 s intervals for ~8 min using a Zeiss LSM 710 laser scanning confocal microscope (objective 60×, 1.3 oil plan-Apochromat; Munich, Germany). Representative time-stamps in (A,B) were extracted from Videos S12 in Presentation 2 and S15 in Presentation 3 of Supplementary Material, respectively, and the time-stamps in (C–E) were from Videos S16–S18 in Presentation 3 of Supplementary Material, respectively. In all time-stamps, target beads are indicated with an arrowhead and in (b; 130 s) a second cell with a pre-captured target bead is indicated with an arrow.

Live-cell imaging of IpLITR 1.1b-mediated target interactions at different incubation temperatures. Rat basophilic leukemia-2H3 cells (3 × 10⁵) stably co-expressing IpLITR 1.1b and LifeAct-GFP were incubated at 37°C (A,B) or at 27°C (C–E) with 9 × 10⁵ αHA monoclonal antibody-coated 4.5 µm bright blue microspheres. Immediately after the addition of target beads, images were collected at 10 s intervals for ~8 min using a Zeiss LSM 710 laser scanning confocal microscope (objective 60×, 1.3 oil plan-Apochromat; Munich, Germany). Representative time-stamps in (A,B) were extracted from Videos S12 in Presentation 2 and S15 in Presentation 3 of Supplementary Material, respectively, and the time-stamps in (C–E) were from Videos S16–S18 in Presentation 3 of Supplementary Material, respectively. In all time-stamps, target beads are indicated with an arrowhead and in (b; 130 s) a second cell with a pre-captured target bead is indicated with an arrow.