Extended Data Fig. 8: Increased numbers of effector T cells in naive mice overexpressing Gch1, and enhanced T cell proliferation after stimulation.

a, Representative immunoblot to detect GCH1 and the HA tag in naive CD4⁺ T cells from control and GOE;Lck-overexpressing mice. The experiment was repeated three times with similar findings. b, c, The proportion of splenic T and B cells (b) and the proportion of CD4⁺ and CD8⁺ T cells among the splenic T cell (TCRβ⁺) population (c), from control (n = 4) and GOE;Lck (n = 4) mice. Data for individual mice aged eight weeks are shown as means ± s.e.m. NS, not significant (two-tailed Student’s t-test). d, Quantification of naive (CD44^lowCD62L^high), memory (CD44^highCD62L^high) and effector (CD44^highCD62L^low) T cell subtypes from the spleen of control (n = 6) and GOE;Lck (n = 10) mice. Data for individual mice are shown as means ± s.e.m. **P < 0.01; ***P < 0.001; NS, not significant (two-tailed Student’s t-test). e, Cell numbers for B cells, T cells, and CD4⁺ and CD8⁺ T cells in the spleens of control and GOE;Cd4 mice. Data from individual mice (n = 4 for each genotype) are shown as means ± s.e.m. NS, not significant (two-tailed Student’s t-test). f, Proportion of CD4⁺ and CD8⁺ naive, memory and effector T cells in the spleens of naive control and GOE;Cd4 mice. Data for individual mice (n = 4 for each genotype) are shown as means ± s.e.m. *P < 0.05; **P < 0.01; ***P < 0.001; NS, not significant (two-tailed Student’s t-test). g, Representative histograms depicting dose-dependent proliferation of control and GOE;Cd4 CD4⁺ T cells stimulated for three days with anti-CD3/28 antibodies. Experiments were repeated more than three times with comparable results. h, IL-2 and IFN-γ secretion after three days of stimulation (with anti-CD3/28 antibodies) of control and GOE;Cd4 CD4⁺ T cells. Data are shown as means ± s.e.m. n = 3 for each genotype. *P < 0.05; ***P < 0.0001 (two-tailed Student’s t-test). i, j, Cells from control (n = 3) and GOE;ERT (n = 3) mice were stimulated with anti-CD3/28 antibodies for three days and treated with 4-hydroxytamoxifen (4-OHT; 0.5 μM) to induce Gch1 overexpression in vitro. i, Quantification of proliferation of CD4⁺ T cells; j, cytokine secretion. Data from individual mice are shown as means ± s.e.m. **P < 0.01; ***P < 0.001 (two-tailed Student’s t-test).