Extended Data Fig. 5: Rescuing altered redox ratio and aspartate level is not sufficient to rescue the stimulation-induced inhibition of nucleotide de novo synthesis.
a) Basal oxygen consumption rate of unstimulated or stimulated BMDMs. Mean ± SD (n = 10 independent samples). b) Intracellular NAD to NADH ratio in unstimulated or stimulated BMDMs with or without supplementation of 2.5 mM pyruvate. c) Relative intracellular abundance of aspartate in BMDMs over a timecourse of continual stimulation. d) Relative intracellular abundance of aspartate in unstimulated or stimulated BMDMs, with or without supplementation of 2.5 mM pyruvate or 5 mM aspartate throughout the timecourse. e–i) Labeled fraction of (e) M + 2 labeled IMP, (f) M + 2 labeled AMP, (g) M + 1 and M + 3 labeled GMP, (h) M + 1 and M + 2 labeled CDP, and (i) M + 1 labeled UMP after 24 h γ-15N-glutamine labeling in unstimulated or stimulated BMDMs, with or without supplementation of 2.5 mM pyruvate or 5 mM aspartate throughout the timecourse. N.d. indicates not detected. a–b, d–i) Stimulated cells are continually stimulated for 48 h. b–i) Mean ± SD (n = 3 independent samples). a) Statistical analysis was performed with unpaired two-tailed student’s t-test comparing unstimulated to stimulated cells. c) Statistical analysis was performed using one-way ANOVA followed by post hoc Dunnett’s test comparing all groups to unstimulated cells (0 h). b, d–i) Statistical analysis was performed using one-way ANOVA followed by post hoc Tukey’s test. Bars with different lower-case letters (a, b, c, d, or e) indicate a statistically significant difference with p < 0.05.
