Mus musculus
Protocols: CD1 male mice (Charles River Laboratories, Germany) age 11 weeks were divided into two groups: vehicle (4 animals) and lipopolysaccharide (LPS)-treated mice (4 animals). The LPS-treated group was given a single intraperitoneal injection of LPS from Escherichia coli O111:B4 (Sigma Aldrich, L2630) at a dose of 5 mg/kg, dissolved in 0.9% NaCl. Vehicle mice were injected with a solution of DMSO/Tween80/NaCl (10%/10%/80%). The mice were sacrificed 6 hours later by anesthetizing the animals with isoflurane followed by decapitation. Brains were quickly frozen and stored at -80oC Nuclei were prepared using the NUC201 isolation kit from Sigma Aldrich. Briefly, 8 x 50$\mu$m sagittal sections of cortex from each animal were prepared using a microtome and placed in 200 $\mu$l of cold Nuclei Pure Lysis Buffer (Nuclei Pure Prep Nuclei isolation kit - Sigma Aldrich) with 1M dithiothreitol (DTT) and 0.2 U $\mu$l−1 SUPERase inhibitor (Invitrogen) freshly added before use. Nuclei were extracted using a glass dounce homogenizer with Teflon pestle using 10-12 up and down strokes in lysis buffer. 360 $\mu$l of cold 1.8M Sucrose Cushion solution was added to lysate which was then filtered through a 30-$\mu$m strainer. 560 $\mu$l of filtered solution was carefully overlayed on 200 $\mu$l of Sucrose solution and nuclei were purified by centrifugation for 45 min at 16,000g. The nuclei pellet was re-suspended in 50 $\mu$l cold Nuclei Pure Storage Buffer (Nuclei Pure Prep Nuclei isolation kit - Sigma Aldrich) with 0.2 U $\mu$l−1 SUPERase inhibitor and centrifuged for 5 min at 500g. The supernatant was removed, the pellet washed again with Nuclei Pure Storage Buffer with 0.2 U $\mu$l−1 SUPERase inhibitor and centrifuged for 5 min at 500g. Finally, the pellet was re-suspended in 50 $\mu$l cold Nuclei Pure Storage Buffer with 0.2 U $\mu$l−1 SUPERase inhibitor. cDNA libraries from each sample were prepared using the Chromium Single Cell 3' Library and Gel Bead kit v3 (10x Genomics) according to the manufacturer's instructions.
ENA-FIRST-PUBLIC
2019-08-11T04:02:19Z
ENA-LAST-UPDATE
2019-07-30T14:26:48Z
External Id
SAMEA5818840
INSDC center name
UZH
INSDC first public
2019-08-11T04:02:19Z
INSDC last update
2019-07-30T14:26:48Z
INSDC status
public
Submitter Id
E-MTAB-8192:LC025
age
11
broker name
ArrayExpress
common name
house mouse
developmental stage
adult
fraction
nuclei
genotype
wild type genotype
individual
LC025
organism part
cerebral cortex
sample name
E-MTAB-8192:LC025
scientific_name
Mus musculus
sex
male
strain
CD-1