Extended Data Fig. 7: The mTORC2 signaling generally inhibits ferroptosis in memory CD4+ T cells.
LM-GP61 or IAV-GP61 is the recombinant Listeria monocytogenes or Influenza A Virus expressing LCMV glycoprotein-specific I-Ab restricted CD4 T cell epitope GP66-77. a, Experimental setup of inducible knockout of Rictor in BMCs generated with BM cells from C57BL/6 (WT, CD45.1+) and ERT2Cre-Rictorfl/fl (Rictor−/−, CD45.2+) mice at day 60 post infection of either LM-GP61 or IAV-GP61. b-e, Bodipy C11 intensity on memory CD4+ T cells in spleens of BMCs after administration of Tamoxifen at day 60 post infection with LM-GP61 (b) or IAV-GP61 (d) as described in a. Ratio between Rictor−/− (CD45.2+) and WT (CD45.1+) viral specific memory CD4+ T cells (GP66-77+) in PBMCs from BMCs before and after administration of Tamoxifen at day 60 post infection with LM-GP61(c) or IAV-GP61 (e) as described in a. The presented data are representative of two independent experiments (n=4 in b-e). f, Experimental setup and gating strategy for flow cytometry. Human naïve (Tnaive), central memory (TCM), and effector memory (TEM) CD4+ T cells isolated from PBMCs of healthy human donors were treated with Torin-1, Rapamycin, or vehicle, then analyzed via flow cytometry. g, h, Bodipy C11 intensity (g) and percentage of dead cells (Live/Dead+) in enumerated CD4+ T cell subsets in PBMCs after treatment of Torin-1, Rapamycin, or vehicle as described in f. The presented data are representative of two independent experiments (n=5 in g-h). Data are analyzed by paired two-tailed t-test in b-e. Data are presented as mean values ± SD and analyzed by unpaired two-tailed t-test in g-h.
