Colon cancer cell lines were grown in arginine free media (Thermofisher, Cat # A2493901, US Bio Cat# D9803–07B) supplemented with 10% dialyzed FBS (ThermoFisher, Cat# 26400044) and with arginine supplementation (Sigma Aldrich Cat #A8094). Lysine (Sigma Aldrich Cat #A8094) and bicarbonate were supplemented to DMEM reference levels (Supplemental table 1 ). Cells were plated into 96 well plates (3000 cells per well) and cell viability was assessed with a luminescence-based assay (CellTiter Glo, Promega, Cat # G7572) at 48 hours on a SpectraMax M3 plate reader (Molecular Devices). For nucleotide rescue experiments, nucleobases were supplemented at concentrations up to 10x reported physiologic concentrations (55 (link)) (Sigma Aldrich Cat #A2786, #C3506, #G11950, #T0895). All cell lines were periodically assessed for mycoplasma contamination by PCR for genomic DNA.
A2786
Manufactured by Merck Group
The A2786 is a laboratory-grade centrifuge designed for high-speed separation of samples. It features a robust construction, digital controls, and variable speed settings to accommodate a wide range of applications.
Automatically generated - may contain errors
2 protocols using a2786
1
Colon Cancer Cell Growth in Arginine-Free Media
2
Allopurinol and Nucleoside Metabolism in Flies
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Allopurinol (A8003, Sigma-Aldrich) was diluted in standard medium at 100 μg/ml and flies were placed for 5 d on this medium before metabolite extraction. Adenine, adenosine, and hypoxanthine (A2786, A9251, and H9377, Sigma-Aldrich) or N6-methyladenosine (m6A) (QB-1055, Combi-Blocks, San Diego, CA) were diluted in fly food medium at 500 μM. Parents were allowed to lay eggs on this medium in order to have exposition to the drug throughout all larval development of the progeny. Adults were collected and placed in normal medium until 5 d before the test, when they were placed again in food supplemented with adenosine or m6A at the same concentrations.
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