Extended Data Fig. 5: Related to HK2 dependence is not linked to a direct gene-nutrient interaction.
(a, c, e, g, j, k) Rates of net exchange for glucose (a), glutamine (c), lactate (e), pyruvate (g), and alanine (j, k) for HK2-knockout cells (mean ± s.e.m., n = 3 biologically independent samples). Two-tailed Welch’s t-test between bars. Blue box (k, bottom right), exogenous alanine levels at 72 h vs 24 h (mean ± s.d., n = 3 biologically independent samples). No exchange (ne) was defined as a change in exogenous levels less than 5%. (b, d, f, h, i) Exogenous levels of glucose (b), glutamine (d), lactate (f), pyruvate (h), and alanine (i) during log growth of HK2-knockout cells (mean ± s.e.m., n = 3 biologically independent samples). Blue box (i, right), exogenous alanine levels at 72 h vs 24 h (mean ± s.d., n = 3 biologically independent samples). (l) Immunoblot for expression of HK1 in WC and IP:HA from HK2-knockout cells that expressed HA-MITO. COXIV served as a mitochondrial control marker. S6K served as a non-mitochondrial control marker. (m) HK1 signal normalized by COXIV signal in IP:HA versus WC from the same respective cells. (n) Fractional labeling of lactate in HK2-knockout cells (mean ± s.d., n = 3 biologically independent samples). Values above brackets indicate differences in fractional labeling between bars. Two-tailed Welch’s t-test. (o, p, q, r) 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. For o, p, and q, metabolites that comprised each respective pool of HPLM-specific metabolites can be found in Fig. 5i.
