PI3K inhibitors have minimal effect on the development of NK cells <i>in vivo</i>.

<p>Wild-type C57BL/6 mice were orally dosed with vehicle, the p110α selective inhibitor MLN1117 (60 mg/kg), or the pan-PI3K inhibitor GDC-0941 (70 mg/kg) for 7 days (n = 4). Spleen and bone marrow (BM) were isolated and analyzed for NK cell development by flow cytometry. (<b>A</b>) Splenocytes were stained with CD21 and CD23 for marginal zone (MZ) B cell population. Among the B220⁺ population, CD21^highCD23^low cells were gated for MZ B cell population. (<b>B, C</b>) Percentage of B/T cells and Th/Tc cells in spleen were determined using surface staining with anti-B220/anti-Thy1.2 mAbs and anti-CD4/anti-CD8 mAbs, respectively. (<b>D</b>) NK cells as specified by CD3⁻NK1.1⁺ were gated and analyzed in single-cell suspension of spleen and BM, respectively. (<b>E</b>) Spleen- or BM-derived CD3⁻NK1.1⁺ NK cells were analyzed for NK subset based on the expression level of CD11b and CD27. (<b>F</b>) Spleen- or BM-derived CD3⁻NCR1⁺ NK cells were analyzed for maturation marker, Ly49A, LY49C/I, Ly49D, Ly49G2, and Ly49H. (<b>G</b>) Spleen- or BM-derived CD3⁻NCR1⁺ NK cells were analyzed for the expression of CD122, NKG2D, and NKG2A/C/E. The data are expressed as the means ± SEM. Statistical analysis was performed with the unpaired Student's <i>t</i> test using Sigma Plot 10 software to compare the differences between vehicle and each inhibitor-treated group. *, <i>p</i><0.05, **, <i>p</i><0.01, and ***, <i>p</i><0.001.