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. Author manuscript; available in PMC: 2012 Oct 15.
Published in final edited form as: J Immunol. 2011 Sep 14;187(8):4151–4160. doi: 10.4049/jimmunol.1003137

FIGURE 1.

FIGURE 1

Preferential downregulation of H2-O following in vivo DC activation by multiple TLR agonists. A, B6 mice were injected intravenously with saline, or the indicated TLR agonist, and 16 hr later spleens were harvested and subjected to flow cytometric analysis. Cells were gated for lack of B220 expression and then plotted as CD11c versus MHCII. DCs were defined as MHCII+CD11C+ as indicated by the black box. DCs were further separated based upon CD8α and DEC205 expression to define CD8α DEC205 (CD8α) and CD8α+DEC205+ (CD8α+) DC subsets as indicated. Histograms show the levels of CD86, H2-M, and H2-O (black lines) for CD8α and CD8α+ DCs. Grey lines on top histograms (Saline) for H2-O and H2-M represent the level of non-specific staining. Numbers on plots for H2-O and H2-M represent the percent of cells falling within the Low and High gates as indicated. Vertical dashed lines on CD86 histograms are provided to aid comparison of CD86 levels. B, Quantification of the CD86 mean fluorescence intensity (MFI) for CD8α and CD8α+ DCs after in vivo DC activation. The MFI for each condition is normalized to that observed for control mice injected with saline to allow for comparisons between experiments. C and D, H2-O and H2-M levels were separated into low and high expressing populations as indicated in A on the histograms for Saline injected control mice and the % of H2-OLow (C)and H2-MLow (D) among the CD8α and CD8α+ DCs after in vivo activation with TLR agonists were quantitated for multiple experiments. E, Ratio of intracellular H2-M to H2-O MFI. Data is compiled from three independent experiments. The statistical significance of potential differences between the percent H2-OLow DCs, the percentage of H2-MLow DCs and the H2-M:H2-O ratio obtained for DCs obtained from saline injected compared to TLR agonist injected mice was determined by the Student’s T test. ns, nonsignificant.