Extended Data Fig. 9: FASN is not required for breast tumor growth in the liver.
From: Fatty acid synthesis is required for breast cancer brain metastasis
a, Heatmap showing complex lipid levels measured by LCMS in extracellular fluid (ECF) isolated from the MFP, brain, and liver of non-tumor bearing NSG mice. Data presented within each row were z-score normalized. The data for Brain and MFP ECF are the same as that shown in Fig. 4d. (Brain ECF, n = 2; MFP ECF, n = 2 ECF samples pooled from up to ~8-10 mice). b, Heatmap showing complex lipid levels measured by LCMS of MFP, brain and liver tissue from non-tumor bearing NSG mice. Data presented within each row were z-score normalized. The data for brain and MFP are the same as that shown in Fig. 4e. (Brain tissue, n = 2; MFP tissue, n = 2; Liver Tissue, n = 2). c, Western blot analysis of acetyl-CoA carboxylase (ACC1), ATP citrate lyase (ACLY), fatty acid synthase (FASN), and stearoyl-CoA desaturase-1 (SCD1) in BT474 brain and liver tumor tissue. β-actin expression was assessed as a loading control, and relative densitometry values (normalized to β-actin expression) were used for quantitation and are presented below each blot. Differences in protein expression between brain and liver BT474 lesions was compared by two-tailed t-test. d, The percent of fully labeled (m + 6) glucose in blood plasma following a 12 h 30 mg/kg/min 13C-glucose infusion into female Nude mice bearing BT474 lesions in the liver as assessed GCMS (n = 5 plasma from mice bearing BT474 liver tumors). e, The distribution of pyruvate labeling in BT474 tumors growing in the liver and in non-cancerous liver tissue was measured by GCMS following a 12 h 13C-glucose infusion into female Nude mice. Each isotopologue was normalized to 13C-glucose labeling in plasma (Liver tumor, n = 6; Liver tissue, n = 5). f, The distribution of 13C-label in even isotopologues of palmitate derived from saponified lipids in BT474 liver tumors and in non-cancerous liver tissue following a 12 h 13C-glucose infusion into female Nude mice was measured by GCMS and normalized 13C-glucose labeling in plasma (Liver tumor, n = 6; Liver tissue, n = 5). g, Tumor growth over time of liver tumors in NSG mice generated from control and FASN-disrupted (sgFASN_1) BT474 cells (presented in Fig. 5a-g). Tumor volumes were measured by ultrasound. (BT474 control tumors, n = 4; BT474 sgFASN_1 tumors, n = 4). MAG-monoglyceride; DAG-diglyceride; TAG-triglyceride; Cer-ceramide; LPC- lysophosphatidyl-choline; LPE-lysophosphotidyethanolamine; PC-phosphatidylcholine; PE-phosphatidylethanolamine; PI-phosphatidylinositol; PS-phosphatidylserine; SM-sphingomyelin; CE-cholesteryl ester; @-sphingosine, palmitoylethanolamide, 7−Dehydrodesmosterol, cholesterol, coenzyme Q9, coenzyme Q10. Data presented in panels d, e, f and g are means ± s.e.m.
