characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2022-09-03T00:15:59Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2022-09-03T00:15:59Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA110644859
characteristics.characteristic: External Id
characteristics.text: Suzhou Ninth People's Hospital
characteristics.characteristic: INSDC center name
characteristics.text: 2022-09-03T00:15:59Z
characteristics.characteristic: INSDC first public
characteristics.text: 2022-09-03T00:15:59Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS12742570
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-12079:G4_input_2
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: primary cell line
characteristics.characteristic: cell_line
characteristics.text: mesenchymal stem cell
characteristics.characteristic: cell_type
characteristics.text: premolars extracted for orthodontics
characteristics.characteristic: clinical information
characteristics.text: human
characteristics.characteristic: common name
characteristics.text: Protocols: GMSCs were separated from the surface of the middle third of the root, and were digested with α-minimum essential medium (α-MEM, Gibco, Carlsbad, CA, USA) containing 3 mg/mL collagenase (type I) and 4 mg/mL dispase (both from Sigma-Aldrich, Billerica, MA, USA) for 15 min at 37 °C. Single-cell suspensions were obtained by passing the digested tissues through a 70-μm cell strainer (BD Bioscience, Franklin Lakes, NJ, USA). Then the cell suspensions were seeded into 10-cm culture dishes (Corning, USA) containing α-MEM supplemented with 15% fetal bovine serum (FBS, Hyclone, New York City, NY, USA), and 1 x GlutaMAX (Thermo Fisher Scientific, Waltham, MA, USA). The cells were incubated at 37 °C in a humidified atmosphere containing 5% CO2, and medium was changed twice a week. Cells were used at passages 3-5 for subsequent experiments. SP600125 (MedChemExpress LLC, Shanghai, China) were dissolved in dimethyl sulfoxide (DMSO). Unless otherwise indicated, cells were pre-incubated with serum-free medium containing same dose of DMSO (0.2%) or 10 μM SP600125 for 2 h Genomic DNA was sonicated into 200 bp around fragments, and incubated with the antibody of H3K27me3 (A2363, Abclonal) at 4 oC overnight. This was followed by incubating for 2 h with Protein A beads (Thermo Fisher Scientific) at 37 °C to pull down the bound DNA fragments. We then isolated the DNA fragments from the pull-down by ethanol precipitation. For high throughput sequencing, we added 3’-dA overhang to the H3K27me3 enriched or input DNA and ligated them to the adapter to build a DNA library.
characteristics.characteristic: description
characteristics.text: adult
characteristics.characteristic: developmental stage
characteristics.text: wild type genotype
characteristics.characteristic: genotype
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: gingiva
characteristics.characteristic: organism part
characteristics.text: Homo sapiens
characteristics.characteristic: scientific_name
characteristics.text: G4_input_2
characteristics.characteristic: title