Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2022 Oct 29;13:6459. doi: 10.1038/s41467-022-34156-1

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Fig. 4 — a Cartoons depicting putative mechanisms of trogocytosis via intraluminal vesicle (ILV) generation or membrane fusion between donor and recipient cells. b CD80 experiences different pH-levels upon trogocytosis via CD28 versus CTLA4. CD28- or CTLA4-expressing 58αβ A2 T cells were co-cultured with CD80-TagRFP-pHluorin expressing CHO cells. Quantification of pHluorin/TagRFP ratio and extrapolation of pH-levels experienced by trogocytosed CD80 based on standard curve shown in Figure S3. c, d Analysis of membrane fusion by bi-molecular fluorescence complementation (BIFC) assay. c Cartoon depicting putative outcomes of BIFC trogocytosis assay. CD8+ T cells (d) or CD28- or CTLA4-expressing 58αβ A2 T cells (e) expressing cytosolic GFP1–10 were co-cultured with CD80-mScarlet-GFP11-expressing CHO cells. Positive control: GFP1–10 transgenic T cells transduced with CD80-mScarlet-GFP11 (d). Negative control: GFP1–10 transgenic T cells cultured with CD80-mScarlet-transgenic CHO cells. Plot shows increase of GFP gMFI over negative control after 24 h (e). f Fluorescent tag attached to CD80 cytoplasmic domain is cleavable by extracellular protease treatment. CD28-expressing 58αβ A2 T cells having trogocytosed CD80-IgG-mScarlet were treated with or without trypsin. Negative control: T cells cultured without CHO cells. Representative flow cytometry plots depicting loss of trogocytosed mScarlet and cell surface CD80 signal upon trypsinization-treatment. Note that loss of both signals marks the extracellular location of the cytoplasmic domain of trogocytosed CD80. See also Figures S3 and S4. Statistics: b: pHluorin/TagRFP: pooled data from 13 independent experiments. Two-sided Wilcoxon matched-pairs signed rank test (n = 13 biologically independent samples per group) pH: two-sided paired t test, pooled data from 3 independent experiments (n = 3 biologically independent samples per group). 4e: Pooled data from 3 independent experiments. Two-way repeated-measure ANOVA with Šídák’s correction, sample vs control (n = 6 biologically independent samples per group). Source data are provided as a Source data file.