Extended Data Fig. 1: Characterization of H9T-expanded CD8+ T cells. | Nature

Extended Data Fig. 1: Characterization of H9T-expanded CD8+ T cells.

From: An engineered IL-2 partial agonist promotes CD8+ T cell stemness

Extended Data Fig. 1

a, Pre-activated mouse CD8+ T cells were rested overnight and cultured with PBS, IL-2, H9, or H9T as indicated for 10 min, and western blotted for phospho-STAT5 (pSTAT5) and pERK. Total STAT5 and ERK were included as controls (on the same gel of pSTAT5 and pERK). Data are representative of two independent experiments. Relative densitometry is shown below each panel (normalized to the IL-2 condition). For gel source data, see Supplementary Fig. 1b. b, Pre-activated CD8+ T cells were rested overnight and stimulated with 10 nM IL-2 or H9T, lysed, and immunoprecipitated with anti-STAT5A or anti-STAT5B antibodies followed with western blotting analysis for pSTAT5 and STAT5A or STAT5B as loading controls (on the same gel). Data are representative of two independent experiments. Relative densitometry is shown below each panel (normalized to the IL-2 condition). For gel source data, see Supplementary Fig. 1c. Data are from two independent repeats. c, Pre-activated CD8+ T cells were rested overnight, cultured with PBS, IL-2, H9, or H9T for 6 days, fixed, permeabilized and intracellularly stained for pSTAT5; n = 4 mice. Data are presented as mean values ± SEM, one-way ANOVA test with Dunnett’s correction. Data are from two independent repeats. d, Pre-activated mouse CD8+ T cells were rested overnight and then cultured in medium containing serial dilutions of IL-2, H9, or H9T for 8 days and analysed for TIM-3 expression. Data are representative of three independent experiments. eh, Pre-activated CD8+ T cells were cultured with 10 nM IL-2, H9, or H9T for 8 days, and surface expression of TIM-3 (e), PD-1 (f), LAG-3 (g), and 2B4 (h) was analysed by flow cytometry. Data are representative of four independent experiments

Source data.

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