2024-01-01
ENA first public
ERC000011
ENA-CHECKLIST
IBCH RAS
INSDC center name
public
INSDC status
ERS24379932
SRA accession
E-MTAB-15106:U87_TMZ2
Submitter Id
ArrayExpress
broker name
U87 MG
cell line
Glioblastoma cell line
cell type
not collected
collection date
human
common name
Protocols: Cells were seeded in 25 mm² culture flasks and treated with temozolomide for 5 days. Senescent cells were subsequently cultured for 24 hours without the drug. Control cells were grown in 25 mm² culture flasks and harvested at 80% confluency. Cell were cultivated in DMEM (PanEco, Russia) supplemented with 10% fetal bovine serum (FBS) (HyClone, USA) at 37°C and 5% CO2, and dissociated by EDTA solution (PanEco, Russia).  To induce senescence, the cells were pre-treated with temozolomide (TMZ) (Macklin, China) for at least 5 days. TMZ concentration was pre-selected to be sub-toxic (50 µM). Total RNA was extracted from cell culture lysates using the Triazol reagent and the PureLink RNA Micro Kit (Invitrogen) according to the protocol provided with the reagent. The quality and quantity of the extracted total RNA were assessed using the BioAnalyzer instrument and the RNA 6000 Nano Kit (Agilent). Subsequently, the total RNA was used to obtain the ribominus fraction using the NEBNext® rRNA Depletion Kit v2 (Human/Mouse/Rat) (NEB, E7400L) following the manufacturer's instructions. Sequencing libraries were prepared from the ribominus RNA fraction using the NEBNext® Ultra™ II RNA Library Prep Kit (E7770S, NEB) according to the manufacturer's protocol. The concentration of the libraries was determined using the Qubit dsDNA HS Assay Kit (Thermo Fisher Scientific) on a Qubit 2.0 instrument. The size distribution of the library fragments was assessed using the Agilent High Sensitivity DNA Kit (Agilent). Sequencing was performed on the HiSeq 1500 platform (Illumina), generating at least 40 million short reads with a length of 150 nucleotides each.
description
adult
developmental stage
brain glioblastoma
disease
PTEN-null, TP53 wild-type
genotype
not collected
geographic location (country and/or sea)
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
brain
organism part
Homo sapiens
scientific_name
U87_TMZ2
title