Expression of IL-4 and IL-5 by lung CD4⁺ cells in <i>Lilrb4^+/+</i> and <i>Lilrb4</i>^−/− mice.

<p>Mice were sensitized and challenged with OVA as described in the legend for <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0057007#pone-0057007-g002" target="_blank">Fig. 2</a>. After 18 h, lung mononuclear cells were obtained as described in the legend for <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0057007#pone-0057007-g001" target="_blank">Fig. 1</a>, and cells were analyzed by intracellular flow cytometry to determine the percentage, number, and MFI of IL-4 and IL-5 in CD4<b>⁺</b> cells. Data from representative experiments show dot plots for IL-4 and IL-5 (A); positivity was defined as cells that had fluorescence intensities greater than that of 99% of the same population of cells when stained with an equal amount of isotype control mAbs. Compiled data for IL-4 (B) and IL-5 (C) are expressed as mean ± SEM, <i>n</i> = 8.</p

Expression of CCR7 on lung DCs in <i>Lilrb4^+/+</i> and <i>Lilrb4</i>^−/− mice.

<p>Mice were sensitized with OVA/LPS and challenged with AF-OVA as described in the legend for <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0057007#pone-0057007-g001" target="_blank">Fig. 1</a>. After 4 h, lung mononuclear cells were obtained as described in the legend for <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0057007#pone-0057007-g001" target="_blank">Fig. 1</a>, and CD11c^+/autofluorescence<b>⁻</b> cells were analyzed by flow cytometry for the percentage (<i>A</i>) and MFI (<i>B</i>) of CCR7 expression on AF-OVA<b>⁺</b> and AF-OVA<b>⁻</b> DCs. Fluorescence compensation was set for each color such that there was no cross-talk between detection channels. Data are expressed as mean ± SEM, <i>n</i> = 5.</p

Expression of CCL21 in the lung lymphatics of <i>Lilrb4^+/+</i> and <i>Lilrb4</i>^−/− mice.

<p>Animals were sensitized with OVA and LPS (<i>A</i>; left and middle panels) or sham sensitized with PBS (<i>A</i>; right panel) as described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0057007#pone-0057007-g001" target="_blank">Fig. 1</a> legend on days 0, 1, and 2. Mice were challenged with 25 µg of unlabeled OVA intranasally on day 14. After 4 h, mice were euthanized, and paraffin-embedded sections of lungs were prepared. Tissue sections were incubated with rabbit anti-mouse LYVE-1 and goat IgG anti-mouse CCL21. Tissue sections were washed and incubated with Alexa Fluor 488-labeled donkey anti-goat IgG (green), Alexa Fluor 594-labeled chicken anti-rabbit IgG (red), and nuclear dye Hoechst 33342 (blue). Sections were washed, mounted, and examined by fluorescence microscopy. Representative photomicrographs (<i>A</i>) and compiled data (<i>B</i>) are presented. Data in <i>B</i> are expressed as mean ± SEM; <i>n</i> = 9–10.</p