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

. 2017 Apr 3;214(4):1181–1198. doi: 10.1084/jem.20161263

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

© 2017 Inoue 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 5. — Decreased antigen presentation of Foxo1-deficient LZ GC B cells. (A, left) Flow cytometry of surface molecules on Foxo1^+/+ and Foxo1^f/f LZ GC B cells. Mice were immunized as described in Fig. 2 A, and LZ GC B cells were prepared on day 12. (right) Histograms indicating the gMFI of each population. n = 3 biological replicates. (B) In vitro antigen presentation assay. Mice were prepared as described in Fig. 3 A. Purified splenic B cells were incubated with NP-Eα-GFP or Eα-GFP for 1 h at 37°C. Antigen-presenting LZ GC B cells were detected by staining the LZ GC B cells (CD45.1⁺CD138⁻GL7⁺CD38⁻CD86^hiCXCR4^lo) with Y-Ae antibody. (left) Flow cytometry of LZ GC B cells; (middle) the percentage of Y-Ae⁺ (bracketed line in the left) LZ GC B cells; (right) the gMFI of Y-Ae of Y-Ae⁺ LZ GC B cells. n = 3 biological replicates. (C) In vitro antigen presentation of Foxo1-deficient LZ GC B cells using αDEC-Eα-GFP. Eα-GFP and αDEC-OVA were used as negative controls. Mice were prepared as described in Fig. 3 A. (left) Flow cytometry of LZ GC B cells; (right) the percentage of Y-Ae⁺ (bracketed line in the left) LZ GC B cells. n = 3 biological replicates. (D) Effects of Foxo1 ablation in LZ GC B cells on T_FH cell number and cytokine production. (left) Schematic representation of the experimental protocol. Foxo1^+/+ ERT2cre B1-8^hi CD45.1⁺CD45.2⁺ or Foxo1^f/f ERT2cre B1-8^hi CD45.1⁺CD45.2⁺ B cells were co-transferred together with CD4⁺ T cells from OT-II CD45.1⁺CD45.2⁺ mice into wild-type mice (CD45.1⁻CD45.2⁺), which were immunized with NP-OVA/alum i.p. on day 0. Mice were administered tamoxifen p.o. on day 7, 8, and 9, and analyzed on day 12. (middle) Flow cytometry of OT-II CD4⁺ T cells (CD45.1⁺CD4⁺). (right) The percentage of T_FH cells (CD45.1⁺CD4⁺CXCR5⁺PD-1⁺) among total donor OT-II cells, and the quantification of Il21 mRNA expression in sorted T_FH cells by real-time qPCR. n = 8 biological replicates. Error bars represent SD. Data are representative of three (A) or two (B–D) independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001; unpaired Student’s t test.