Figure 6.

Defective NK cell activation in the absence of mTOR.

(a-d) CD45.1 Mtor^WT/WT or CD45.2 Mtor^lox/lox splenocytes treated in vitro with TATCre were transferred into CD45.1/CD45.2 double positive hosts. 2 days later, mice were injected or not with poly(I:C). Splenic cell suspension were analyzed 18 h later by flow cytometry. (a) Histograms represent phosphorylation of S6 and Akt proteins (one representative experiment out of 3 (t-test, *p<0.05, **p<0.01, ***p<0.001, ns = non significant). independent experiments using 1 mouse in each), numbers indicate the MFI. The scatter plot shows phosphoproteins MFI in NK cells from poly(I:C) injected mice (n=5 mice, 3 independent experiments). MFI is normalized to the resting population. (b) The scatter plot shows FSC MFI in NK cells from poly(I:C) injected mice (n=6 mice, 3 independent experiments). (c) Histograms represent expression of CD71, CD98 and CD69 or the amount of 2-NBDG staining in one experiment out of 3, numbers indicate the MFI. The scatter plot shows MFI in NK cells from poly(I:C) injected mice (n=6 mice, 3 independent experiments). (d) Histograms show intracellular expression of GzmB in one representative experiment out of 3. The scatter plot shows MFI normalized to the resting population (n=5 mice, 3 independent experiments). (a-d) In all scatter plots, lines link the CD45.1 Mtor^WT/WT and CD45.2 Mtor^−/− NK cells co-transferred in the same host. (e) NK-Mtor^WT/WT and NK-Mtor^−/− mice were infected with MCMV and sacrificed at day 0, 2 or 6.5. Expression of intracellular Ki67, GzmB and IFN-γ analyzed in Ly49H⁺ and Ly49H⁻ splenic NK cell subsets. The number of Ki67⁺ NK cells as well as the GzmB MFI (normalized to non-infected mice) and the percentage of IFN-γ⁺ NK cells in each subset is shown (average +/− SD, dots represent individual mice, statistical comparisons have been made between NK-Mtor^WT/WT and NK-Mtor^−/− NK cells for each time point). Ki67⁺ NK cell proliferation is indicated. Results are from more than at least 6 mice, 4 independent experiments. (t-test, *p<0.05, **p<0.01, ns = non significant).