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. Author manuscript; available in PMC: 2021 Oct 8.
Published in final edited form as: Cell Rep. 2021 Sep 14;36(11):109701. doi: 10.1016/j.celrep.2021.109701

Figure 5. NaCT supports extracellular citrate import and metabolism in hepatocellular carcinoma cells.

Figure 5.

(A) Citrate uptake flux in Huh7 NTC and SLC13A5 KO cells grown in hypoxia for 48 h (n = 3).

(B) Citrate transport in Huh7 NTC and SLC13A5 KO #1 cells (n = 3).

(C) Mole percent enrichment of TCA intermediates from [2,4-13C2]citrate in Huh7 NTC and SLC13A5 KO cells grown in hypoxia for 48 h (n = 3).

(D) Mole percent enrichment of palmitate from [2,4-13C2]citrate in Huh7 NTC and SLC13A5 KO cells grown in hypoxia for 48 h (n = 3).

(E) Mole percent enrichment of citrate from [U-13C5]glutamine in Huh7 NTC and SLC13A5 KO cells ± 500 μM citrate grown in hypoxia for 48 h (n = 3).

(F) Percent labeling of M3 aspartate from [U-13C5]glutamine in Huh7 NTC and SLC13A5 KO cells ± 500 μM citrate grown in hypoxia for 48 h (n = 3).

(G) Percentage of lipogenic acetyl-CoA contributed by [U-13C5]glutamine in Huh7 NTC and SLC13A5 KO cells ± 500 μM citrate grown in hypoxia for 48 h (n = 3).

α-KG, α-ketoglutarate; Suc, succinate; Mal, malate; Asp, aspartate. In all graphs data are plotted as means ± SD. Statistical significance is relative to NTC as determined by one-way ANOVA with Dunnett’s method for multiple comparisons (A, C, and D) or relative to (−) citrate as determined by two-sided Student’s t test (B, E, and F) with *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. In (G), data are plotted as mean ± 95% CI. Asterisk (*) indicates statistical significance by non-overlapping CIs. Unless indicated, all data represent biological triplicates. See also Figures S3 and S4. (C–G) Data shown are from one of at least two separate experiments.