Sb415286

Dulbecco’s modified Eagle medium (DMEM) and fetal bovine serum (FBS) were purchased from Gibco (Carlsbad, CA, USA). LY294002, SB415286, DCFH-DA and MPP⁺ were purchased from Sigma Chemicals (St. Louis, MO, USA). PD98059, Anti-phospho-p44/42 MAPK (ERK1/2, Thr202/Tyr204), anti-ERK, anti-phospho-GSK3 β (Ser9), anti-GSK3β, anti-phospho-Akt (Ser473), anti-Akt, anti-phospho-JNK, anti-JNK, anti-phospho-p38, anti-p38 and anti-phospho-CREB (Ser133) were obtained from Cell Signaling Technology (Boston, MA, USA). The antibody against GAPDH and secondary antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

3(4,5-dimethylthiazol-2-yl)-2.5-diphenyltetrazolium bromide, LY294002, Hoechst 33342, SB415286, DCFH-DA, FDA, PI and L-glutamic acid hydrochloride (glutamate) were from Sigma Chemicals (St. Louis, MO, United States). Cell culture medium and fetal bovine serum were from Gibco (Carlsbad, CA, United States). Phospho-Ser473-Akt, total Akt, phospho-Ser9-GSK3β, total GSK3β and β-actin antibodies were from Cell Signaling Technology (Beverly, MA, United States).

Cycloheximide, actinomycin D, MG132, insulin, LiCl, SB415286 and DAPI were purchased from Sigma-Aldrich. The restriction endonucleases (NheI, XhoI, XbaI, EcoRI, KpnI and BamHI) and recombinant GSK-3β were bought from New England Biolabs. All of the primers used were synthesized from Integrated DNA Technologies. Protein A/G plus agarose was purchased from Santa Cruz. The anti-HA, anti-Myc antibodies and SignalSilence Control siRNA, SignalSilence GSK-3α/β siRNA were obtained from Cell Signaling; anti-actin was obtained from Sigma-Aldrich; anti-SCF^Fbw7 was obtained from Abcam and anti-SREBP-1 was bought from Becton-Dickinson and Co. GenJet Plus transfection reagent was purchased from SignaGen Laboratories and Halt™ combined protease and phosphatase inhibitor cocktails were bought from Thermo Scientific. Trypsin and Lys-C enzymes for mass spectrometry and the Dual-Luciferase® Reporter (DLR™) Assay System were obtained from Promega. SCF^Fbw7 siRNA 1 (s30664), SCF^Fbw7 siRNA 2 (s224357), SimplyBlue and Lipofectamine RNAiMAX were purchased from Invitrogen. AdEasy XL Adenoviral Vector System was purchased from Agilent Technologies.

Fucoxanthin was extracted from Sargussum horneri using a series of steps, including solvent extraction, ethanol precipitation and low-temperature concentration. Briefly, fucoxanthin isolation was conducted at 30°C for 2 h with ethanol to sample ratio of 4:1 (v/w). Then the fucoxanthin-containing solution was concentrated at 25°C. Lipid and chlorophylls were precipitated when the ethanol content reached 63% in the concentrated solution. Fucoxanthin was purified by precipitation when the ethanol content reached 40% in the solution. The purity of fucoxanthin was more than 90% by HPLC, and purified fucoxanthin was stored at −20°C prior to further analysis. H₂O₂ was purchased from Calbiochem (San Diego, CA, USA). SB415286 was purchased from Sigma Chemicals (St Louis, MO, USA). U0126, Wortmannin and LY294002 were supplied from LC Laboratories (Woburn, MA, USA). Antibodies against pSer473-Akt, Akt, pSer9-GSK3β, GSK3β, pERK and ERK were provided by Cell Signaling Technology (Beverly, MA, USA). Unless otherwise noted, all media and supplements for cell cultures were obtained from Invitrogen (Carlsbad, CA, USA).