Extended Data Fig. 2: Normal T cell development and B cell biology in the absence of Gch1.

a, Thymocyte cell death induced over 24 h by various stimuli: anti-CD3 (0.5 μg ml⁻¹ and 5 μg ml⁻¹), Fas ligand (0.2 μg ml⁻¹ and 2 μg ml⁻¹), dexamethasone (Dex, 0.1 μg ml⁻¹ and 0.5 μg ml⁻¹) and γ-irradiation (1 Gray (Gy)). Data are shown as means ± s.e.m. n = 3 for each genotype. NS, not significant (two-tailed Student’s t-test). b, Death by neglect of purified CD4⁺ T cells cultured without stimulation for up to 56 h. Data are shown as means ± s.e.m. n = 3 for each genotype. NS, not significant (two-tailed Student’s t-test). c, d, Proliferation of CD4⁺ T cells from control and Gch1;RORc mice after three days of anti-CD3/28 stimulation. Panels show representative FACS proliferation traces (c) and representative dose response (d). Experiments were repeated independently more than six times with similar results. e, Representative FACS plots from spleens of control and Gch1;MB1 mice. MB1-Cre is an early B cell deleter Cre line using endogenous CD79a B cell specific expression. The experiment was repeated two independent times with similar results. f, g, Representative FACS histogram depicting the proliferation of wild-type B cells treated with vehicle (DMSO) or SPRi3 (50 μM) (f), and of B cells from control and Gch1;MB1 mice in response to LPS (1 μg ml⁻¹) after three days (f). Shaded grey peaks represent unstimulated cells. FACS plots are representative of two independent experiments showing similar results. n = 3 mice per group. h, Class-switch recombination. FACS analysis of splenic CD43⁻ B cells from control and Gch1;MB1 mice stimulated with LPS (20 μg ml⁻¹) for five days to induce class-switch recombination to IgG3. FACS plots are representative of two independent experiments showing similar results.