Glutamine carbon is associated with glutamine nitrogen in building pyrimidine rings in PCa. A, Schematic overview of the synthesis of pyrimidine. B and C, Schematic overview of glutamine carbon flux toward related metabolites. Gln, glutamine. Asp, aspartate. OAA, oxaloacetic acid. α-KG, α-ketoglutarate. Ac-CoA, acetyl-CoA. PC, pyruvate carboxylase. GLS1, glutaminase. GOT, glutamic oxaloacetic transaminase. RC, reductive carboxylation. ADT, androgen deprivation therapy. D-I, Fractions of 13C5-glutamine derived isotopologues in aspartate (D and G), UTP (E and H) and CTP (F and I) in the indicated PCa cell lines. J-L, Fractions of 13C5-glutamine derived isotopologues in aspartate (J), UTP (K) and CTP (L) in LNCaP cells treated in regular medium or charcoal-stripped medium (ADT). Data are depicted as mean ± s.d. * P < 0.05, ** P < 0.01 and *** P < 0.001 by two-tailed Student’s t-test.