Figure 3.

mTOR activity is primarily under IL-15 control.

(a) Splenocytes were cultured for 1 h on plates coated with the indicated antagonistic antibodies or with the indicated cytokine. Cells were subsequently stained and the averaged MFI (+/− s.d.) of intracellular pS6 in NK cells is shown (n=4 independent experiments using 1 mouse each time). (b) Splenocytes were cultured with graded concentrations of IL-15 for 1 h. Cells were stained and the averaged MFI of intracellular pS6 and pSTAT5 in NK cells are given as percentage of the maximal response (+/− s.d.) (n=3 independent experiments using 1 mouse each time). (c) Freshly isolated BM cells of control or anti-CD122 injected mice were stained and analyzed by flow cytometry. The amount of pS6 as a function of CD27 expression in one representative experiment out of 3 is shown. The bar graph shows averaged phosphoproteins MFI of BM NK cells from control of anti-CD122 treated mice (n=3 mice in 3 independent experiments). MFI is normalized to the CD11b^lo subset. (d) Histograms represent phosphorylation of different proteins in splenic NK cells from control or poly(I:C) injected mice 4 h before, with or without anti-CD122, one representative experiment out of 3 is shown. The bar graph shows averaged phosphoproteins MFI (+/− s.d.) (average of n=3 mice in 3 independent experiments). Fold change (FC) in MFI is obtained by normalizing the activated to the resting population. (t-test, *p<0.05, **p<0.01, ***p<0.001, ns = non significant).