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Series GSE236324 Query DataSets for GSE236324
Status Public on Jun 13, 2024
Title Enhanced crosslinking and immunoprecipitation analysis of human fetal β-cells
Organism Homo sapiens
Experiment type Other
Summary N6-methyladenosine (m6A) is the most abundant chemical modification in mRNA, and plays important roles in human embryonic stem cell pluripotency, maintenance, and differentiation. However, the role of m6A and the precise mechanisms involved during the development of β-cells are unexplored. Here, we performed enhanced crosslinking and immunoprecipitation (eCLIP) assays to identify m6A sites regulated by METTL14 in EndoC-βh1 human fetal β-cells.
 
Overall design EndoC-βH1 cell lines were obtained from Univercell-Biosolutions (France). Culture plates were coated with DMEM (glucose 4.5 g L-1; Gibco) containing fibronectin (2 μg mL-1; Gibco), and extracellular matrix (1% vol vol-1; Sigma) for at least an hour in 5% CO2 at 37°C. EndoC-βH1 cells were grown on coated 6-well plates containing DMEM (glucose 1 g L-1), BSA fraction V (2% wt vol-1) (Roche), 2-mercaptoethanol (50 μM; Sigma), nicotinamide (10 mM; Sigma), transferrin (5.5 μg mL-1; Sigma), and sodium selenite (6.7 ng mL-1; Sigma). The eCLIP assays were performed by Eclipse Bioinnovations using UV-crosslinked EndoC-βH1 cells following the protocol detailed in (Van Nostrand et al. Nat Methods. 2016) using the anti-METTL14 antibody A305-847A (Bethyl Laboratories).
 
Contributor(s) Kahraman S, DeJesus DF, Kulkarni R
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Submission date Jul 03, 2023
Last update date Jun 14, 2024
Contact name Hui Pan
E-mail(s) Hui.Pan@joslin.harvard.edu
Organization name Joslin Diabetes Center
Department Bioinformatics and Biostatistics Core
Street address 1 Joslin Place
City Boston
ZIP/Postal code 02215
Country USA
 
Platforms (1)
GPL16791 Illumina HiSeq 2500 (Homo sapiens)
Samples (4)
GSM7527973 METTL14-n1-input
GSM7527974 METTL14-n1-IP
GSM7527975 METTL14-n2-input
Relations
BioProject PRJNA990571

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Supplementary file Size Download File type/resource
GSE236324_RAW.tar 143.6 Mb (http)(custom) TAR (of BED, BIGWIG)
SRA Run SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file

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