Human normal pancreatic duct cell line hTERT-HPNE (CRL-4023; ATCC, Manassas, Virginia, USA) was grown in Dulbecco’s modified Eagle’s medium (E600010; Sangon Biotech, Shanghai, China) with 5% fetal bovine serum (FBS, C0234; Beyotime, Shanghai, China), 10 ng/mL recombinant human EGF (P5552; Beyotime), 5.5 mM d -glucose (G7021; Sigma-Aldrich), and 750 ng/mL puromycin (ST551; Beyotime) as recommended by the producer. PCa cell lines BxPC3 (C1023), SW1990 (C1196), and PANC1 (C1004) were available from WHELAB (Shanghai, China). BxPC3 cells were maintained in RPMI-1640 medium (E600028; Sangon Biotech), SW1990 cells were cultured in F-12 basic medium (M0500; WHELAB), and PANC1 cells (C1004; WHELAB) were grown in the high-glucose DMEM (M0100; WHELAB). All media for PCa cells were blended with 10% FBS and 1% penicillin–streptomycin (G0100; WHELAB). Cell incubation was completed in an incubator (HF100; Heal Force, Shanghai, China) at 37°C with 5% CO2.
D glucose g7021
Manufactured by Merck Group
Sourced in United States
D-glucose (G7021) is a monosaccharide that serves as a primary source of energy in many biological processes. It is a common laboratory reagent used in various applications.
Automatically generated - may contain errors
Lab products found in correlation
Htert hpne, by American Type Culture Collection (1 mentions)
Hf100, by Heal Force (1 mentions)
Puromycin st551, by Beyotime (1 mentions)
E600028, by Sangon (1 mentions)
Acetylcholine a6625, by Merck Group (1 mentions)
Tissue tek, by Sakura Finetek (1 mentions)
Accu chek aviva nano, by Merck Group (1 mentions)
Polyvinylidene difluoride pvdf membrane, by Merck Group (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Dmi8 microscope, by Leica (1 mentions)
Giemsa, by Merck Group (1 mentions)
Fibronectin, by Merck Group (1 mentions)
M1902, by Merck Group (1 mentions)
Polydimethylsiloxane pdms, by Dow (1 mentions)
Neutrophil isolation kit, by STEMCELL (1 mentions)
Fmlp f3506, by Merck Group (1 mentions)
6 protocols using d glucose g7021
1
Culture and Maintenance of Pancreatic Cell Lines
2
Tissue Preparation and Compound Acquisition
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Ellagic acid (E2250), Dihydroethidium (DHE) methyl-beta-cyclodextrin (332615), acetylcholine (A6625) and D-glucose (G7021) were obtained from Sigma-Aldrich (St. Louis, MO, USA). The optimum cutting temperature compound (OCT), Tissue-Tek, was obtained from Sakura Finetek (Torrance, CA, USA).
3
Fluorescent Membrane Permeability Assay
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Cell membrane impermeable β GREEN-np (Spectragenetics) reconstituted in HBSS, the working concentration is 100 nM for imaging, 200 nM for FACS. Botulinum Neurotoxin Type C1 was purchased from List Biological Laboratories. Adenosine 5′-triphosphate magnesium salt (A9187), EDTA and D-(+)-Glucose (G7021) were obtained from Sigma.
4
Metabolic Phenotyping in Mice
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Assessment of metabolic parameters was conducted at 4 months of age within 2 consecutive weeks according to the standard operating procedures of the National Institutes of Health (NIH) Mouse Metabolic Phenotyping Center [30 (link)]. For the glucose tolerance test, animals were fasted overnight for 15 h (±1 h), starting at 6:00PM with free access to water. Blood glucose was measured before and 15, 30, 60, and 120 min following an intraperitoneal injection of 2 g/kg of a 20% glucose solution (d -glucose, G7021, Sigma Aldrich), with one drop of blood from the tail-tip using a glucometer (Accu-Chek Aviva Nano). For the insulin tolerance test, animals were fasted for 6 h (±1 h), starting at 9:00AM with free access to water. Blood glucose was measured before and 15, 30, 60, and 120 min following an intraperitoneal injection of 1 IU/kg insulin (Insulin solution, I9278, Sigma Aldrich), with one drop of blood from the tail-tip using a glucometer (Accu-Chek Aviva Nano). The area under the curves (AUCs) for the tolerance tests were calculated using the trapezoidal rule (GraphPad Prism, version 8). For the baseline blood glucose levels, blood glucose levels were measured after an overnight fasting of 15 h (±1 h), with one drop of blood from the tail-tip using a glucometer (Accu-Chek Aviva Nano).
5
Antibodies and Reagents for Cell Assays
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Primary and secondary antibodies used in this study are shown in Supplemental Table 1 . Polyvinylidene difluoride (PVDF) membranes were purchased from Millipore (Darmstadt, Germany). The drugs used were as follows: streptozotozin (STZ, S0130); RPMI 1640 medium without glucose (R1383); (+)-JQ1 (BRD4 inhibitor, SML1524); and D-glucose (G7021), all purchased from Sigma-Aldrich (St Louis, MO, USA); and D-Luciferin potassium salt (C3654) purchased from APExBIO (Houston, TX, USA).
6
Neutrophil Isolation and FITC-Dextran Assay
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Neutrophil isolation kit (17957, STEMCELL, Vancouver, Canada); Bovine serum albumin (A1933-25G, Sigma-Aldrich, St. Louis, MO, USA), FBS (iCell-0500, iCell Bioscience Inc, Shanghai, China); RPMI 1640 (SH30809.01, Hyclone, Logan, UT, USA); Fluorescein (FITC-dextran, 10 kDa; final concentration 5 μM, Sigma-Aldrich, St. Louis, MO, USA); fMLP (F3506, 1 mM, Sigma-Aldrich, St. Louis, MO, USA); Triton X-100 (9036-19-5, Sigma, St. Louis, MO, MO, USA); Giemsa (G5637, Sigma, St. Louis, MO, USA); D-glucose (G7021, Sigma-Aldrich, St. Louis, MO, USA); mannitol (10 mM, an osmotic control reagent, M1902, Sigma-Aldrich, St. Louis, MO, USA); Fibronectin (F2006, 2 μg mL−1, Sigma-Aldrich, St. Louis, MO, USA); Commercial magnetic Neutrophil isolation kit (17957, STEMCELL Technologies, Inc., Toronto, Canada); Precursors and initiators of Polydimethylsiloxane (PDMS) (Dow Corning, Midland, MI, USA); Glass slide (80350-0001, Jiangsu Shitai Xinchuang Scientific Instrument Co., Ltd., Nanjing, China); Microscope (DMi8, Leica, Wezler, Hesse, Germany); Plasma cleaner (PDC-002, Harrick Scientific Products, Inc., New York, NY, USA).
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