SLP-76 is important for Ly49 receptor expression and activation downstream of multiple NK cell activating receptors.

<p>(<b>A</b>) Representative histograms of NK cell (gated on NKp46⁺DX5⁺ splenocytes) expression of multiple activating receptors are shown (WT: tint; SLP-76 KO: black line). (<b>B)</b> Representative histograms of CD107a expression by WT and SLP-76 KO NK cells post-stimulation from four independent experiments are shown. (<b>C</b>) %CD107a expressing NK cells are represented as mean ± standard error of the mean (SEM) of four independent experiments (<i>n</i> = 6 mice). Ly49H-stimulated NK cells are gated on Ly49H⁺ cells. (<b>D</b>) The percent and (<b>E</b>) total number of splenic NK cells (NK1.1⁺NKp46⁺ CD3ε⁻) were calculated and represented as mean percent positive ± SEM of <i>n</i> = 4–5 mice/group. (<b>F</b>) qPCR was performed for SLP-76 (<i>lcp2)</i> and GAPDH (housekeeping gene) with RNA from sorted BM NK cells [CD3ε ⁻CD122⁺NK1.1⁻ (pNK), CD3ε ⁻CD122⁺NK1.1⁺DX5⁻ (iNK), CD3ε ⁻NK1.1⁺DX5⁺ (mNK)], splenic CD3ε ⁻NK1.1⁺DX5⁺ (mNK) and splenic T cells (CD3ε ⁺). Data are from three experimental replicates and is graphed as % of control (splenic mNK). *<i>p</i> < 0.05, **<i>p</i> < 0.01, and ***<i>p</i> < 0.001 by paired student’s <i>t</i> test. (<b>G</b>) Splenic and (<b>H)</b> BM NK cells from WT (black bars) and SLP-76 KO (white bars) mice were assessed for expression of Ly49 inhibitory receptors (Ly49A, Ly49G2, Ly49C, Ly49I), Ly49 activating receptors (Ly49D and Ly49H), and NKG2A/C/E. The proportion of receptor-expressing NK cells from multiple mice is represented as mean percent positive ± SEM of <i>n</i> = 4–5 mice/group. (<b>I)</b> CD27 and CD11b expression by WT (black) and SLP-76 KO (white) NK cells is represented as percent positive ± SEM of <i>n</i> = 5–6 mice. *<i>p</i> < 0.05, **<i>p</i> < 0.01, ***<i>p</i> < 0.001, and ns = not significant by student’s <i>t</i> test. See <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1002526#pbio.1002526.s001" target="_blank">S1 Data</a> for raw data.