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

. 2000 Jul 3;192(1):23–30. doi: 10.1084/jem.192.1.23

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

© 2000 The Rockefeller University Press

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 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

PMC Copyright notice

graphic file with name JEM000305.f3a.jpg — B cell activation in vitro. (a) Purified cells were stimulated with LPS from E. coli 0111:B4, E. coli 055:B5, S. minnesota Re595, and synthetic lipid A. After 3 d, cells were pulsed with 1 μCi/well ³H-TdR and harvested, and [³H]thymidine incorporation was counted as described (reference 9). Results were shown as mean values of cpm from triplicate wells with standard errors. (b) Top: purified B cells were stimulated with the following stimuli: the goat anti–mouse IgM F(ab′)₂ Ab (5 μg/ml), the mAb to CD40 (5 μg/ml), and recombinant mouse IL-4 (5 ng/ml). After 3 d, cells were pulsed with 1 μCi/well ³H-TdR, and B cell proliferation was measured as described above. (b) Bottom: purified B cells were stimulated with goat anti–mouse IgM F(ab′)₂ Abs at various concentrations with or without IL-4, and proliferation was measured as described above. (c) Purified B cells were stimulated with lipid A (1 μg/ml) for 1 h (lanes 2 and 5) or 3 h (lanes 3 and 6). Cell lysates were immunoprecipitated with anti-Lyn mAb, and equally divided precipitates were subjected to SDS-PAGE (9% acrylamide) and electroblotting. Phosphotyrosine (top) or Lyn (bottom) was probed with antiphosphotyrosine or anti-Lyn mAb, respectively. Similar results were obtained from two independent experiments.

graphic file with name JEM000305.f3b.jpg — B cell activation in vitro. (a) Purified cells were stimulated with LPS from E. coli 0111:B4, E. coli 055:B5, S. minnesota Re595, and synthetic lipid A. After 3 d, cells were pulsed with 1 μCi/well ³H-TdR and harvested, and [³H]thymidine incorporation was counted as described (reference 9). Results were shown as mean values of cpm from triplicate wells with standard errors. (b) Top: purified B cells were stimulated with the following stimuli: the goat anti–mouse IgM F(ab′)₂ Ab (5 μg/ml), the mAb to CD40 (5 μg/ml), and recombinant mouse IL-4 (5 ng/ml). After 3 d, cells were pulsed with 1 μCi/well ³H-TdR, and B cell proliferation was measured as described above. (b) Bottom: purified B cells were stimulated with goat anti–mouse IgM F(ab′)₂ Abs at various concentrations with or without IL-4, and proliferation was measured as described above. (c) Purified B cells were stimulated with lipid A (1 μg/ml) for 1 h (lanes 2 and 5) or 3 h (lanes 3 and 6). Cell lysates were immunoprecipitated with anti-Lyn mAb, and equally divided precipitates were subjected to SDS-PAGE (9% acrylamide) and electroblotting. Phosphotyrosine (top) or Lyn (bottom) was probed with antiphosphotyrosine or anti-Lyn mAb, respectively. Similar results were obtained from two independent experiments.