Chromatin immunoprecipitation-sequencing was conducted as described in (5 (link)), by incubating Beta-TC6 cells (ATCC; CRL-11506) cultured in DMEM supplemented with 15% FBS, 1% L-glutamine, and 1% penicillin/streptomycin with DEC1 (Novus Biologicals; NB100–1800) and DEC2-V5 (GeneCopoeia, Inc. # EX-A4343-LX304) antibodies. Sequencing reads were mapped using STAR (46 (link)) with default parameters. Regions enriched for uniquely-mapped reads relative to input sample were called as binding peaks using MACS (47 (link)) with default parameters and “--pvalue=1e-9 --keep-dup=1”, followed by filtering against overlap with problematic regions of the genome (48 (link)). Peaks from BMAL1, CLOCK, and PDX1 ChIP-sequencing in Beta-TC6 cells (10 (link)) were called in the same manner and concatenated and merged with DEC1 and DEC2 peaks using BEDTools (49 (link)) to obtain a unified peak catalog.
Beta tc6 cells
Manufactured by American Type Culture Collection
The Beta-TC6 cells are a clonal cell line derived from transgenic mice that carry the simian virus 40 (SV40) T antigen gene under the control of the insulin promoter. These cells exhibit characteristics of pancreatic beta cells and can be used for in vitro studies related to beta cell function and diabetes research.
Automatically generated - may contain errors
2 protocols using beta tc6 cells
1
ChIP-seq analysis of DEC1 and DEC2 in Beta-TC6 cells
2
Beta-TC-6 Cells Culture and hIAPP Treatment
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Beta-TC-6 cells (ATCC, Manassas, VA) were maintained in monolayer cultures in HEPES-buffered Dulbecco’s modified Eagle’s medium (Life Technologies, Grand Island, NY) supplemented with 15% heat-inactivated fetal bovine serum (Life Technologies), 100 IU/ml penicillin and 100 μg/ml streptomycin (Life Technologies). Cells were maintained at 37°C humidified atmosphere with 5% CO2. h-IAPP (Bachem, King of Prussia, PA) was dissolved in water and immediately added it to the culture medium of β-cells at various concentrations.
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