Extended Data Fig. 6: Related to HK detachment from the OMM drives glycolytic ATP production.
(a) Relative growth of HK2-knockout versus control cells (mean ± s.d., n = 3 biologically independent samples). Two-tailed Welch’s t-test comparing the respective mean ± s.d. (bar) versus mean ± s.d. (control cells) between bars. (b, c, d, e, f) Relative levels of the indicated metabolites in HK2-knockout versus control cells (mean ± s.d., n = 3 biologically independent samples). Two-tailed Welch’s t-test comparing the respective mean ± s.d. (bar) versus mean ± s.d. (control cells) between bars. Hypoxanthine (Hx), adenosine (Ado), and guanosine (Guo) can serve as salvage substrates for purine synthesis. Cytidine (Cyt) and uridine (Urd) can serve as salvage substrates for pyrimidine synthesis. Thymidine (Thy) is a salvage substrate for thymidylate (b, right). N-acetylglucosamine (GlcNAc) can act as a salvage substrate for UDP-GlcNAc (c, right). Glycerol-3-phosphate (Glycerol-3P) and acetyl-CoA are key intermediates involved in palmitate synthesis (d, right). Dimethyl α-KG (DM-α-KG) may deliver an additional carbon source to the TCA cycle (e, right). NADPH is a key product of the oxidative pentose phosphate (ox-PPP) pathway (f, right). (g) Pyruvate-to-lactate ratios in HK2-knockout and control cells (mean ± s.d., n = 3 biologically independent samples). Two-tailed Welch’s t-test. (h, i) Relative growth of HK2-knockout and control cells treated with duroquinone (DQ) (h) or UK5099 (i) versus vehicle-treated control cells (mean ± s.d., n = 3 biologically independent samples). Two-tailed Welch’s t-test comparing the respective mean ± s.d. (bar) versus mean ± s.d. (control cells) between bars. DQ can serve as an electron acceptor for the NAD+-generating reaction catalyzed by NAD(P)H dehydrogenase [quinone] 1 (NQO). DHQ, durohydroquinone. (h, right). UK5099 is a small molecule inhibitor of the mitochondrial pyruvate carrier (MPC) (i, right). (j) Oxygen consumption rates (OCR) and proton efflux rates (PER) for HK2-knockout and control cells (mean ± s.e.m., n = 6 biologically independent replicates). OA, oligomycin A; Rot, rotenone; AA, antimycin A. (k) Basal OCR (left) and OCR coupled to ATP production (right) for HK2-knockout versus control cells (mean ± s.d., n = 6 biologically independent replicates). Two-tailed Welch’s t-test comparing the respective mean ± s.d. (bar) versus mean ± s.d. (control cells). (l) Total population doublings for control cells (mean ± s.d., n = 3 biologically independent samples). Value above bracket indicates difference in total doublings.
