Figure 5. Klrk1^-/- and Tyrobp^-/- NK cells have reduced levels of CD3ζ and Zap70 signaling molecules.

(a-b) Splenic NK cells from C57BL/6J (n=5 mice per group), Klrk1^-/- (n=4-5 mice per group) and Tyrobp^-/-(n=4 or 6 mice per group) mice were analyzed for expression of CD3ζ, FcεRγ, Zap-70 and Syk. Shown are graphs of geometric mean values for indicated molecules (a) as well as histograms (b). As a controls for staining, isotype control (for CD3ζ and Zap-70), FMO (for FcεRγ) or secondary antibody only (for Syk) were used. FACS plots were gated for NK cells (CD3^-NK1.1⁺). (c) NK cells from C57BL/6J and Klrk1^-/- mice were sorted and expression of indicated proteins was analyzed by western blot. Pan-akt or β-actin were used as controls for equal loading. (d) NK cells from Ncr1^Cre (n=6), Ncr1^CreKlrk1^fl/fl (n=4), Klrk1^-/-(n=5) and C57BL/6J (n=5) mice were analyzed for CD3ζ and Zap-70 expression. Figures show graphs of geometric mean. (e) Transcript levels for CD3ζ and FcεRγ of NK cells sorted from spleens from C57BL/6J and Klrk1^-/- mice were analyzed by qPCR (n=5 mice per group). Shown are representative plots of three (a and c) or two (b and d) experiments. ANOVA, with Bonferroni's post-test correction for multiple comparisons was used to analyze a and c. Two-tailed unpaired t-test was used to analyze d. Shown are means ± s.e.m. ** P<0.01, *** P < 0.001.