Dulbecco s modified eagle s medium high glucose dmem hg

Dulbecco’s modified Eagle’s medium high glucose (DMEM-HG), fetal bovine serum (FBS), and trypsin-EDTA (0.05%) were obtained from Thermo Fisher Scientific. Normal human gastric mucosal epithelial cell lines (GES-1) and GC cell lines (MFC) were acquired from the American Type Culture Collection (Manassas, VA, USA). The cells were incubated in DMEM supplemented with 10% FBS under standard culture conditions (5% CO2, 37 °C).

Copper(II) chloride dehydrate (CuCl₂·2H₂O), hydrochloric acid (HCl, 37%), nitric acid (HNO₃, 65%), and Dulbecco’s modified Eagle’s medium-high glucose (DMEM/HG) were purchased from Thermo Fisher Scientific (Massachusetts, USA). Iron(II) chloride anhydrous (FeCl₂, 99.5%), methylene blue (C₁₆H₁₈ClN₃) and thiazolyl blue tetrazolium bromide (MTT) were from Alfa Aesar (Massachusetts, USA). Hydrazine hydrate (N₂H₄·H₂O) and sodium pyruvate (C₃H₃NaO₃S·xH₂O) were from Acros Organics (Morris Plains, NJ, USA). poly(styrene-alt-maleic acid) sodium salt solution (PSMA), hydrogen peroxide (H₂O₂), 2′,7′-dichlorofluorescein diacetate (DCFH-DA), sodium bicarbonate (NaHCO₃), trypan blue (0.4%), imidazole, N,N-dimethyl-4-nitrosoaniline (RNO), N-(3-dimethylaminopropyl)-N’-ethylcarbodiimide hydrochloride (EDC), N-hydroxysuccinimide (NHS), and folic acid (C₁₉H₁₉N₇O₆) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Fetal bovine serum (FBS), trypsin-ethylenediaminetetraacetic acid(trypsin-EDTA, 0.25%) and antibiotic-antimycotic (penicillin/streptomycin/amphotericin B) were from Biological Industries (Cromwell, CT, USA). Dimethyl sulfoxide was from Scharlau (Barcelona, Spain).

Dulbecco’s modified Eagle’s medium/high glucose (DMEM/HG), fetal bovine serum (FBS), penicillin–streptomycin, and newborn calf serum were purchased from Thermo Fisher Scientific (Waltham, MA, USA), while 3-(4,5 Dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT), isopropanol, dimethyl sulfoxide (DMSO), and 3-isobutyl-1-methylxanthine (IBMX) were obtained from Merck Millipore (Burlington, MA, USA). Dexamethasone, 10% formalin, phosphate-buffered saline (PBS), insulin, and Oil-Red-O were purchased from Sigma-Aldrich (St. Louis, MO, USA). Rosiglitazone and 2-deoxy-2-((7-nitro-2,1,3-benzoxadiazol-4-yl)amino)-d-glucose (2-NBDG) were purchased from Cayman Chemical (Ann Arbor, MI, USA). Antibodies against β-actin, Akt, phospho-Akt (Ser473), AMPKα, and phospho-AMPKα (Thr172) were purchased from Cell Signaling Technology (Danvers, MA, USA). Peroxidase-conjugated goat anti-rabbit and goat anti-mouse IgG were obtained from Merck Millipore.

Oil palm empty fruit bunches (OPEFBs) were gathered from a palm oil mill namely Tennamaram, Selangor, Malaysia. Pluronic F127 (PF127) (molecular weight (MW): 12,600 g/mol), dimethylsulfoxide (DMSO), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), 1-chloro-2,4-dinitrobenzene (DNCB), and sodium lauryl sulfate (SLS) were supplied by Sigma–Aldrich, Hamburg, Germany. SSD was supplied by the Tokyo Chemical Industry, Tokyo, Japan. Human epidermal keratinocyte (HaCaT) cells were purchased from the American Type Culture Collection (ATCC), Virginia, USA. Fetal bovine serum (FBS), penicillin/streptomycin antibiotic, and Dulbecco′s modified Eagle′s medium high glucose (DMEM-HG) were supplied from Thermo Fisher Scientific, Waltham, MA, USA. Ammonia solution (25 v/v%) was supplied by Guangdong Guanghua Sci-Tech Co. Ltd., Shantou, China. Staphylococcus aureus (S. aureus) (ATCC^® 6538) was purchased from Microbiologics Inc., Saint Cloud, MN, USA. Nutrient broth and nutrient agar were obtained from HiMedia Laboratories Pvt. Ltd., Ambernath, India. Normal saline was supplied by B. Braun Medical Inc., Melsungen, Germany. Ethanol was purchased from Systerm chemicals, Shah Alam, Malaysia. Finally, sodium dodecyl sulphate (SDS) was provided by the Promega Corporation, Madison, WI, USA. All chemicals were American Chemical Society (ACS) grade and were used as received.

The purified sulfated polysaccharide (AdSP, polysaccharide content of ~99%, Mw = 272 kDa) extracted from self-grown Arthrospira maxima through the pressurized hot water extraction method was provided by Far East Bio-Tec. Co., Ltd. (Taipei, Taiwan). The molar ratio of each monosaccharide composition is listed in Table 2. Methyl acrylic anhydride (MAA), gelatin (type A, 300 Bloom, from porcine skin), deuterium oxide (D₂O), and sodium bicarbonate were purchased from Sigma-Aldrich (St. Louis, MO, USA). High-glucose Dulbecco’s Modified Eagle’s Medium (HG-DMEM), Ham’s F-12, fetal bovine serum (FBS), and penicillin–streptomycin–amphotericin (PSA) were purchased from Gibco (Grand Island, NY, USA). Geneticin (G418) was purchased from Invitrogen (Waltham, MA, USA). 2,2-Azobis(2-methyl-N-(2-hydroxyethyl) propionamide) (VA-086) and FG were purchased from Wako Chemicals GmbH (Neuss, Germany). Chitosan (Mn~1.4 × 10⁵ Da) was obtained from Hopax lnc. (Taipei, Taiwan). CCK-8 (Sigma-Aldrich, St. Louis, MO, USA) and KAPA SYBR Green qPCR kit (Kapa Biosystems, Inc., Wilmington, MA, USA) were applied in the commercial form.

High-glucose Dulbecco’s modified Eagle’s medium (HG-DMEM), fetal bovine serum (FBS), penicillin, and streptomycin were ordered from Gibco. The EnzChek Phosphatase Assay Kit (catalog no. E12020) and DiFMUP (catalog no. D6567) were purchased from Thermo Fisher Scientific Inc. The Malachite Green Phosphate Detection Kit (catalog no. DY996) was obtained from R&D Systems. Peptide Src pY530 (sequence: STEPQpY QPGENL) was synthesized by ChinaPeptides Co. Ltd.
The following antibodies were used for Western blot experiments: rabbit anti-p44/42 mitogen-activated protein kinase (MAPK) (ERK1/2; Cell Signaling Technology, catalog no. 4695), rabbit anti-phosphothreonine (202)/tyrosine (204)-p44/42 MAPK (pERK1/2; Cell Signaling Technology, catalog no. 4370), rabbit anti-Src (Cell Signaling Technology, catalog no. 2109), rabbit anti–hemagglutinin (HA) (Cell Signaling Technology, catalog no. 3724), rabbit anti-Flag (Cell Signaling Technology, catalog no. 14793), rabbit anti-Src (phospho Y529/530) (Abcam, catalog no. ab32078), rabbit anti–glyceraldehyde phosphate dehydrogenase (Beyotime Biotechnology, catalog no. AF5009), and goat anti-rabbit IgG/horseradish peroxidase (HRP) (Beyotime Biotechnology, catalog no. A0208).