Extended Data Figure 5. FBP1 regulates glycolysis, glutamine metabolism, and pentose phosphate pathway (PPP) in renal cells.

Glucose uptake (a) and lactate secretion (b) in HK-2 cells with or without FBP1 inhibition, cultured in medium containing 1 mM glucose. c, Carbon fate map showing the isotopomer distribution of indicated metabolites derived from [1, 2-¹³C] glucose. ¹³C atoms are depicted as filled circles. ¹³C atoms directly going through the glycolytic pathway are coloured in black, while ¹³C atoms going through the PPP and recycled back to glycolysis are coloured in red. d, Glutamine uptake in RCC10 cells ectopically expressing vector or FBP1. e, Carbon fate map showing the isotopomer distribution of indicated metabolites derived from [U-¹³C] glutamine. f, M4 isotopomer distribution of indicated metabolites in RCC10 cells expressing vector or FBP1, labelled with [U-¹³C] glutamine. % M4 enrichment represents the mole percent excess of M4 species above natural abundance. g, Fold changes of PPP-related metabolites detected in ccRCC tumours vs. adjacent normal kidney. Note that generation of reduced glutathione (G-SH) requires NADPH, a major reducing product of PPP. p value is calculated based on Welch’s paired t-test. q value is the estimation of false discovery rate in multiple testing. h, Relative NADPH levels in HK-2 cells with or without FBP1 inhibition. i, M1 and M2 isotopomer distribution of lactate in in HK-2 cells with or without FBP1 inhibition. j, Calculated PPP flux (relative to vector control) in HK-2 cells with or without FBP1 inhibition. M1 and M2 isotopomer distribution of lactate (k) and calculated PPP flux (l) in RCC10 cells expressing vector or FBP1. Relative glucose 6-phosphate (G6P) levels in HK-2 cells with or without FBP1 ablation (m), and in RCC10 cells expressing vector or FBP1 (n). o, Western blot analysis of indicated proteins in RCC10 and RCC10VHL cells ectopically expressing vector or FBP1. Experiments were performed in triplicates. Values represent mean±s.d. *p<0.05.