Figure S4.

FAMIN Affects Routing through Central Purine Metabolism, Related to Figure 4

(A) Methylthioadenosine levels in Famin^p.254I, Famin^p.254V, Famin^p.284R M1 macrophages (n = 12).

(B) Fraction of inosine, adenine and hypoxanthine labeled as the indicated isotopomer in M1 macrophages after a 3 h pulse with [¹³C₁₀¹⁵N₅] adenosine (n = 6).

(C) Metabolic fate of stable isotope-labeled [¹⁵N₅] adenine after a 3 h pulse of M1 macrophages (n = 6; mean). Schematic representation of central purine metabolism. AMP can be generated from adenine via sequential FAMIN and ADK activities or via APRT without loss of label (purple). Labeled [¹⁵N₅] adenosine can be deaminated into inosine by FAMIN or ADA, with a loss of a single ¹⁵N as ammonia, generating a [¹⁵N₄] isotopomer (brown). Routes of interconversion and relationship with other metabolic pathways are also illustrated. Fractions of different labeled states averaged across genotypes following the 3 h pulse with [¹⁵N₅] adenine are depicted as pie charts; asterisks indicate metabolites with significantly altered isotopic labeling across genotypes as depicted in Figures S4D and S4E. Adenine phosphoribosyl transferase (APRT); adenosine kinase (ADK); adenosine deaminase (ADA); cytosolic nucleotidase (cN); hypoxanthine-guanine phosphoribosyl transferase (HPRT); purine nucleoside phosphorylase (PNP); S-methyl-5′-thioadenosine phosphorylase (MTAP); S-adenosylhomocysteine hydrolase (SAHase).

(D) [¹⁵N₅] adenine levels in M1 macrophages after a 3 h pulse with [¹⁵N₅] adenine (n = 6).

(E) Fraction of adenosine labeled as the [¹⁵N₅] isotopomer in M1 macrophages after a 3 h pulse with [¹⁵N₅] adenine (n = 6).

(F) Fraction of ATP, NAD⁺, NADH, FAD, acetyl-CoA, HMG-CoA and succinyl-CoA labeled as the indicated isotopomer in M1 macrophages after a 3 h pulse with [¹³C₁₀¹⁵N₅] adenosine. Fractions of different labeled states (averaged across Famin^p.254I, Famin^p.254V and Famin^p.284R genotypes) are depicted as pie charts. Asterisks indicate significantly altered isotopic labeling across Famin genotypes (n = 6 per genotype).

Data are represented as mean ± SEM. ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001 (unpaired, two-tailed Student’s t test or one-way ANOVA).