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. 2023 Sep 22;8(18):e169308. doi: 10.1172/jci.insight.169308

Figure 3. The polyamine/hypusine axis suppresses CD69 expression in activated mouse CD8+ T cells.

Figure 3

(A) Schematic of the polyamine-hypusine pathway and selected pharmacologic inhibitors. (B) CD69 MFI in polyclonal CD8+ T cells activated in replete RPMI-1640 medium (with 2 mM glutamine) without (Ctrl) or with added 5 mM DFMO ± 500 μM putrescine (Put), 100 μM spermidine (Spd), or 100 μM spermine (Spm) 72 hours after activation (n = 3). (C) Breeding scheme for the generation of the CD4-Cre+/–;Odcfl/fl mice. (D) Fold change in levels of Odc mRNA (determined by qPCR) normalized to B2m mRNA in CD8+ T cells from CD4-Cre+/–;Odcfl/fl mice (n = 3). (E) CD69 MFI in CD8+ CD4-Cre+/–;Odcfl/fl T cells ± 500 μM Put (n = 3). (F and G) CD69 MFI in purified polyclonal CD8+ T cells (F), or CD8+ OT-I T cells (G), activated under control (Ctrl) conditions or with added 5 mM DFMO ± Put or 10 μM GC7 ± Put at 72 hours after activation (n = 3). Data in B and E–G were analyzed by 1-way ANOVA with Tukey’s post hoc test, and data in D were analyzed using unpaired t test. Each dot represents a biological replicate, and all data are shown as mean ± SEM. **P < 0.01; ***P < 0.001; ****P < 0.0001.