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

. 2014 Jun 2;211(6):1231–1242. doi: 10.1084/jem.20131853

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

© 2014 Ali 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 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

PMC Copyright notice

Figure 4. — TLR priming increases GBS suppression of Sig-14^−/− but not Sig-14^+/+ neutrophil responses. (A) Human neutrophils of the indicated genotypes were left unstimulated or stimulated with LPS for 6 h, incubated with H2DFCA for 30 min, and then infected with GBS (MOI = 10); ROS production was measured 20 min after infection by FACS. The histogram shows fluorescence of ROS indicator H2DCFDA. (B) Human neutrophils of the indicated genotypes were left unstimulated or stimulated with LPS for 6 h and infected with GBS (MOI = 10), and NET formation was visualized by myeloperoxidase and DAPI staining 30 min after infection. Bar, 50 µm. (C) Quantification of NETs shown in B. (D) Human neutrophils of the indicated genotypes were left unstimulated or stimulated with LPS for 8 h; expression of Siglec-5 and Siglec-14 was analyzed by FACS using antibody recognizing both human Siglec-5 and Siglec-14. The histogram depicts combined surface expression of Siglec-5 and Siglec-14. (E) Human neutrophils of the indicated genotypes were stimulated or not with LPS for 6 h and infected with GBS (MOI = 10), and bacterial killing was assayed 20 min after infection. (F) Human neutrophils of the indicated genotypes were stimulated with LPS for 6 h and infected with GBS (MOI = 10), and cell lysates prepared at the indicated times were immunoprecipitated with Siglec-5– and Siglec-14–recognizing antibody. SHP-1 recruitment and total IgG were analyzed by immunoblot. The SHP-1/IgG (×10) densitometry value is shown on the immunoblot. Data in this figure are representative of two to four independent experiments with one to three different donors per genotype in each experiment. Results are means ± SD; *, P < 0.05, **, P < 0.01.