Decanoic acid

Bovine serum albumin (BSA) was purchased from Equitech-Bio, Inc. (Kerrville, TX, USA). Decanoic acid, sodium hydroxide, Hoechst 33258 (H), and fluorescein (F) were purchased from Sigma–Aldrich (Milwaukee, WI, USA). Coumarin 540A (C) was purchased from the Exciton Chemical Co. Inc. (Dayton, OH, USA). Rhodamine B (R) was purchased from the Eastman Kodak Company (Rochester, NY, USA).

Decanoic acid (purity ≥ 98.0%) was purchased from Sigma-Aldrich (St. Louis, MO, USA). HGF was obtained from PeproTech (Offenbach, Germany). Antibodies of Cyclin D1, β-actin, IAP-1, Akt, PARP, Bcl-2, COX-2, and Survivin were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies of ERK, MEK, PI3K, c-Met, p-ERK, p-MEK, p-AKT, p-PI3K, and cleaved caspase-3 were procured from Cell Signaling Technology (Beverly, MA, USA).

Dihydroconiferyl alcohol (DCA, 98%), octanoic acid (≥98%), decanoic acid (≥98%), lauric acid (for synthesis), tert-butanol (≥99%), propyl gallate (PG, ≥98%), ethyl acetate (≥99.5%), acetonitrile (≥99.9%), 2-Chloro-4,4,5,5-tetramethyl-1,3,2-dioxaphospholane (TMDP), N-Hydroxy-5-norbornene-2,3-dicarboxylic acid imide (NHND, 97%), 5% Pd/C and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals were purchased from Sigma-Aldrich. (Hoeilaart, Belgium). Immobilized lipase B from C. antarctica (Novozym^® 435) was a gift from Novozymes (Bagsvaerd, Denmark). Molecular sieves (3 Å, 1.6–2.5 mm, technical, water absorption capacity 0.2 g/g) were acquired at VWR. Hexanoic acid (98%) and silica gel 60 (42–60 µm, 230–400 mesh) were purchased from Merck (Hoeilaart, Belgium). Petroleum ether (b.p = 40–60 °C), butylated hydroxytoluene (BHT, 99.8%) and butylated hydroxyanisole (BHA, 98%) were purchased from Acros Organics (Geel, Belgium).
Technical organosolv lignins were supplied by CIMV (botanical origin: wheat straw) and Fraunhofer (botanical origin: beech wood). These lignins were depolymerized as described in Section 2.6.

The lipids in VAT and SAT were extracted using the Bligh and Dyer method23 (link). The collected adipose tissue (300 mg) was mixed with chloroform/methanol/dH₂O (1:2:1 v/v) and sonicated six times for 10 s (each time). The chloroform–lipid layer was then dried under N₂. The extracted lipids were methylated with a methylation kit (Nacalai Tesque, Kyoto, Japan). A methyl heptadecanoate solution (20 μl of 0.02 g/300 μl toluene) was added as an internal standard. Gas chromatography coupled with a flame ionization detector was used to measure the fatty acid methyl esters (GC-2010, Shimadzu Co., Ltd., Kyoto, Japan). The injector port and detector were set to 250 °C. The oven temperature was programmed to increase from 90 °C to 240 °C at 5 °C/min. The peak area of each lipid species was normalized with respect to the internal standard. The amount of fatty acids was calculated from a gas chromatography calibration curve of standard fatty acids (decanoic acid, palmitic acid, oleic acid, and linoleic acid), which were purchased from Sigma-Aldrich (St. Louis, MO, USA) and used without further purification.

Prior to Gas Chromatography-Mass Spectrometry (GC-MS) analysis, active fractions and commercial decanoic acid, undecanoic acid, dodecanoic acid, tridecanoic acid, and tetradecanoic acid (purity > 98%; Sigma–Aldrich, St. Louis, MI, USA) were methylated and ethylated. Samples (30 mg) were diluted with 10 mL acidified methanol or ethanol (1% sulfuric acid) and stirred for 12 h at room temperature. The product of the reaction was suspended in an aqueous solution of 20% Na₂CO₃ (20 mL) and extracted with CHCl₃ (3×, 20 mL, each one) and the solvent was eliminated under vacuum until dry. These derivatives were stored at 4 °C until use in assays [60 (link)].