Extended Data Fig. 1: IL-7 signaling sustains mTORC2 activity in memory CD4⁺ T cells.

a, Naïve SMARTA cells (CD45.1⁺) were adoptively transferred into C57BL/6J recipient mice (CD45.2⁺) and followed by infection with LCMV Arm⁺ the next day. SMARTA cells were analyzed via flow cytometry at D0 (Naïve, green), D5 (early Effector, blue), and D60 (Memory, red) post-infection. CD127 levels on transferred SMARTA cells in the spleens of recipients, from two independent experiments (n=4 for Naive and D5, n=5 for D90). b, p-AKT_Ser473 levels on memory transferred SMARTA cells as in a after stimulation with (red) or without (blue) recombinant human IL-7 ex vivo, from two independent experiments (n=4). c Experimental setup of IL-7 signaling blockade with αIL-7-mAb at the maintenance stage of CD4⁺ T cell memory. d, p-STAT5_Tyr694 and p-AKT_Ser473 levels on memory SMARTA cells in the spleens of recipients as in c after administration with αIL-7-mAb at day 60 post-infection. The presented data are representative of two independent experiments (n=4 for αIL-7-mAb, n=5 for Control). e, f, Validation of the knockout of Rictor. Immunoblot analysis (e) of lysates of sorted memory CD4⁺ T cells from either CD4^Cre-Rictor^fl/fl (Rictor^−/−) or Rictor^fl/fl mice. The presented data are representative of four independent experiments. (f) p-AKT_Ser473 and p-S6_Ser235/236 levels on memory CD4⁺ T cells in the spleens of recipients as in c. The presented data are representative of four independent experiments (n=4). Grey area in histogram indicates staining with rabbit isotype IgG. Data are presented as mean values ± SD and in a-b, d, f. Box plots show min to max, each data point represents one result from an independent experiment, and horizontal bars denote the median in e. Data are analyzed by unpaired two-tailed t-test in a-b, d-f.

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