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

. 2018 Apr 2;215(4):1245–1265. doi: 10.1084/jem.20162042

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

© 2018 Carpier et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms/). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/).

PMC Copyright notice

Figure 4. — Rab6 and Syntaxin-16 control LAT microcluster formation at the immune synapse and LAT phosphorylation. (A) TIRFM images of endogenous LAT in Jurkat cells expressing shC or Rab6-specific shRNA (sh6, sh7), incubated for 10 min on coverslips coated with poly-l-lysine alone (resting conditions) or anti-CD3ε+antiCD28 Abs (αCD3αCD28, activating conditions) before fixation and staining. (B) Quantification, in the evanescent field, of the density of LAT microclusters in Jurkat cells or (C) CD3ζ-GFP in Jurkat cells expressing CD3ζ-GFP. (D) Immunoblot analysis of phospho-LAT, phospho-CD3ζ, total LAT, and Rab6 in Jurkat cells expressing the different shRNA and activated for different times with anti-CD3ε+antiCD28 Abs. (E) Quantification of phospho-LAT and phospho-CD3ζ intensities, normalized on loading control, and expressed as fold increase of intensity at time 0. (F) TIRFM images of LAT in Jurkat cells expressing shC or Syntaxin-16–specific shRNA (sh1, sh5). (G–J) Quantification of the density of microclusters of LAT (G), ZAP70 (H), phospho-LAT (I), and phospho-ZAP-70 (J) in Jurkat cells, expressing control and Syntaxin-16–specific shRNA, activated for 10 min on coverslips coated with anti-CD3ε+antiCD28 Abs. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 (one-way ANOVA). Median is represented by horizontal lines except in E, where mean is presented. All data are from two independent experiments, except for A, B and G, three independent experiments, and in E, three independent experiments each represented by one color. Bars, 5 µm.