Extended Data Fig. 6: D42 FASN KO cell generation and re-expression of WT and mutant enzymes.
From: Snapshots of acyl carrier protein shuttling in human fatty acid synthase
a, Western blot analysis of FASN levels in FASN KO cells, and re-expressed WT and mutant human FASN enzymes, with actin blotted as a loading control (representative of n = 2 experiments). The MW marker in the FASN section is 240 kDa and the corresponding marker in the actin blot is 42 kDa. For blot source data, see Supplementary Figs. 4 and 5. b, Comparison of palmitate synthesis from [U-13C]-glucose in D42 parental cells and the FASN KO clone. The average of n = 3 replicates is plotted with error bars representing the SD. c, Relative abundance of palmitate measured by GC-MS. Data are normalized to heptadecanoic acid as the internal standard and μg protein, and the average abundance of the WT cells set to 1. Each biological replicate (n = 3) is shown, with the mean plotted and error bars for the SD. d, Isotopologue analysis of heavy carbon atoms incorporated into palmitate in each cell line, with the mean of n = 3 replicates shown and error bars of the SD. Isotopologue enrichment was calculated using FluxFix.science.
