2024-08-13
ENA first public
ERC000011
ENA-CHECKLIST
IFOM ETS - The AIRC Institute of Molecular Oncology
INSDC center name
public
INSDC status
ERS18458655
SRA accession
E-MTAB-13914:Gpr126-iECKO1
Submitter Id
18
day
age
ArrayExpress
broker name
not collected
collection date
house mouse
common name
Protocols: WT and Gpr126iECKO ECs were derived from brains of pups at post-natal stage P18 upon tamoxifen treatment. Briefly, the brains were digested enzymatically in combination with gentle dissociation and adult brain dissociation kit [130-107-677]; Miltenyi Biotech). After dissociation, the myelin cell debris and erythrocytes were removed according to the manufacturer protocol. ECs were enriched by depletion of CD45-positive cells with CD45 microbeads (30-052-301; Miltenyi Biotech). After AN2 re-expression according to the manufacturer protocol, pericytes and astrocytes were depleted using AN2 microbeads (130-097-171; Miltenyi Biotech) and ACSA-2 microbeads (130-097-678; Miltenyi Biotech), respectively, followed by positive selection using CD31 microbeads (30-097-418; Miltenyi Biotech).  Total RNA was extracted from fBECs of WT and Gpr126iECKO mice at P18 using Maxwell RSC simplyRNA tissue kits with the Maxwell RSC Instruments (Promega Corporation). The amount of RNA was measured using the Nanodrop technique and its integrity assessed using Agilent Bioanalyzer 2100 with Nano RNA kits (RIN > 8). Libraries for RNA sequencing were prepared following the manufacturer protocols. mRNA-seq indexed library preparation was performed starting from 500 ng of total RNA with the Illumina ligation stranded mRNA kit according to the manufacturer instructions. Indexed libraries were quality controlled on an Agilent Bioanalyzer 2100 with High Sensitivity DNA kits, quantified with Qubit HS DNA, normalized and pooled to perform multiplexed sequencing runs.
description
mouse postnatal
developmental stage
Cdh5-CreERT2/Gpr126flox/flox (Gpr126 KO)
genotype
not collected
geographic location (country and/or sea)
not applicable
isolate
Mus musculus
organism
Mus musculus
organism
brain
organism part
Mus musculus
scientific_name
Gpr126-iECKO1
title