Figure 3.

Metabolomic Analysis under FASN Inhibition and Mechanism of Cell Death

(A) Metabolic pathway from TCA cycle to various lipids. FFA, free fatty acid; FA-CoA, fatty acyl-CoA; LPA, lysophosphatidic acid; PA, phosphatidic acid; DAG, diacylglycerol; PC, phosphatidylcholine; PS, phosphatidylserine; PI, phosphatidylinositol; PS, phosphatidylserine; SM, sphingomyelin; S1P, sphingosine-1-phosphate.

(B) Liquid chromatography-MS of glycerolipids and SLs after 3 h of 6 μM orlistat treatment with or without 50 μM Pal. Data are normalized to controls without orlistat or Pal. Numbers indicate carbon chain lengths followed by degrees of desaturation. One-way ANOVA with Dunnett's test was performed with orlistat (−) Pal (−) as control (n = 3).

(C) Representative image of AP staining of hiPSCs (253G4) after 24 h of orlistat and chicken egg PC treatment.

(D) Quantification of relative AP-positive cell area in hiPSCs (253G4) after 24 h of orlistat and chicken egg PC treatment. Student's t test was performed for each concentration of orlistat (n = 3).

(E) Representative western blot image of cytochrome c subcellular localization in hiPSCs (253G4) 12 h after orlistat treatment. Cyto C, cytochrome c.

(F) Cytosolic fraction of cytochrome c of hiPSCs (253G4) 12 h after orlistat treatment quantified by western blotting. Student's t test, n = 3.

(G) Representative western blotting image of hiPSCs (253G4) 48 h after transfection with either Scr RNA or FASN siRNA. Pal (50 μM) was supplemented 24 h after transfection.

(H) Relative amount of cleaved caspase 3 in hiPSCs (253G4) 48 h after transfection with either Scr RNA or FASN siRNA quantified by western blotting. Pal (50 μM) was supplemented 24 h after transfection. Signal strengths were standardized using those of GAPDH. One-way ANOVA with Dunnett's test was performed with Scr RNA and without Pal as a control (n = 3).

(I) Schematic of metabolic features of undifferentiated hPSCs and consequences of FASN inhibition.

Data are presented as means ± SD; ∗p < 0.05.