Figure 3. Acetate supports lipid synthesis in the absence of ACLY.

A) Measurements of glucose consumption and lactate production (left) and glutamine consumption and glutamate production (right), normalized to cell volume (cell number X mean cell volume), mean +/− SEM of triplicate wells, **, p<0.01; ***, p<0.001. Experiment was performed in glucose-free DMEM + 10% dFBS + 10 mM glucose + 100 μM sodium acetate. B) Experimental design for heavy isotope labeling of fatty acids using [U-¹³C]glucose, with unlabeled acetate present (left) and [1,2-¹³C]acetate, with unlabeled glucose present (right). C) Isotopologue distribution of palmitate after 48 hours labeling in 10 mM [U-¹³C]glucose in Acly^f/f, PC9, PC9-ACLY-WT, PC9-ACLY-H760A MEFs (top). Expressed as percent enrichment of palmitate (bottom), mean +/− SD of triplicates. D) Isotopologues of palmitate after 48 hours labeling in 100 μM [1,2-¹³C]acetate in Acly^f/f, PC9, PC9-ACLY-WT, PC9-Acly H760A MEFs (top). Expressed as percent enrichment of palmitate (bottom), mean +/− SD of triplicates. E) Isotopologues of HMG-CoA upon 6 hours labeling in 10 mM [U-¹³C]glucose (100 μM unlabeled acetate present) in Acly^f/f and PC9 MEFs, mean +/− SEM of triplicates. F) Isotopologues of HMG-CoA upon 6 hours labeling in 100 μM [1,2-¹³C]acetate (10 mM unlabeled glucose present) in Acly^f/f and PC9 MEFs, mean +/− SEM of triplicates. G) Total HMG-CoA quantitation in cells cultured in DMEM + 10% dFBS + 10 mM glucose + 100 μM sodium acetate (unlabeled), mean +/− SEM of triplicates.