Fig. 3

Rapamycin treatment reduced lactate and 2-HG productions in vitro (LC-MS measurements). The relative levels of lactate and 2-HG (a.); the levels of these oncometabolites in nmol/10⁶ cells (b.); the relative levels of succinate, malate and citrate (c.) after 48-h rapamycin treatment (50 ng/mL); The changes in relative level of total lactate (m + 0 and m + 3) were shown after 48-h rapamycin treatment and one-hour U-¹³C-glucose labelling (d.); The reduced level of total 2-HG (m + 0 and m + 1/2/3) was also detected after rapamycin treatment and U-¹³C-glucose labelling (e.) or U-¹³C-glutamine substrate labelling (f.) m + 0 = unlabelled metabolites; m + 3 = 3 ¹³C in lactate from glucose, m + 1/2/3 = 1/2/3 ¹³C in 2-HG from glucose, m + 5 = 5 ¹³C in 2-HG from glutamine, (m + 4 and m + 5 isotopomers were not detectable in these cells after one-hour glucose labelling in 2-HG); Levels of metabolites were expressed relative to the control (control level = 1) and the tables show the absolute levels of labelled and unlabelled metabolites after rapamycin treatment. (n = 3, *:p < 0.05;)