Fig. 5: Let-7 controls CTL differentiation through inhibition of mTOR/ROS pathways. | Nature Communications

Fig. 5: Let-7 controls CTL differentiation through inhibition of mTOR/ROS pathways.

From: Let-7 enhances murine anti-tumor CD8 T cell responses by promoting memory and antagonizing terminal differentiation

Fig. 5

a Gene set enrichment analysis of top hallmark pathways upregulated and downregulated in P14+ naive, 12h-activated and 5-day differentiated CD8 T cells from lin28Tg mice in comparison to those from let-7Tg mice. RNA-seq data are from Figs. 1 and 4. b MFIs of phospho-Erk, phospho-Akt (Thr308), phospho-Akt (S473), phospho-S6 and ROS in P14+ CD8 T cells from WT, lin28Tg or let-7Tg mice activated in vitro (n = 3 per group). c, d Representative FACS analysis of expression of indicated markers in P14+ CTLs from WT, let-7Tg or lin28Tg mice differentiated with rapamycin (rapa) or NAC. e MFIs of indicated proteins and frequencies of CD62L-positive cells presented as a fold change relative to WT. f MFIs of indicated proteins and frequency of CD62L-positive cells in P14+ CTLs from lin28Tg mice differentiated with rapamycin or NAC added at indicated time points. Data are presented as a fold change relative to untreated control. g MFIs of ROS in P14+ CD8 T cells from WT or lin28Tg mice activated with or without rapamycin. h Representative FACS analysis of Tim3 and CD62L expression in P14+ CTLs from lin28Tg mice on Raptor-/- or Rictor-/- background. i, j Experimental design, tumor growth curves (i) and Kaplan–Meier survival curves (j) of WT mice s.c. injected with B16gp33 tumor cells and adoptively transferred with P14+ CTLs from lin28Tg mice differentiated with (n = 8 mice) or without (n = 7 mice) rapamycin+NAC treatment or let-7Tg mice (n = 10 mice). For control group that received no CTLs, n = 8 mice. k Venn diagram showing the number of let-7 target genes in P14+ naive, 12h-activated and 5-day differentiated CD8 T cells from cluster−1 of Fig. 1e and heatmap displaying the expression of these genes. The numbers in c, h indicate the frequency of populations in each quadrant. The numbers in d indicate MFIs. P-values were determined using a two-tailed unpaired Student’s t-test (b, e–g), c (i) and log-rank Mantel-Cox test (j). Data represent three (b–d, h) and two (i, j) independent experiments, or are pooled from three (e, f) and two (g) independent experiments. Source data for b, e–g, i, j are provided as a Source Data file.

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