Mus musculus
Protocols: Mouse-adapted SARS-CoV-2 strain was established in the previous study. Mice were anesthetized and then inoculated viral solution intranasally (i.n.). Viral solutions for inoculation were diluted with PBS and 30 µl of solution contained twice of the 50% lethal dose (2LD50) determined in 24-week-old BALB/c mice. At three post infection, mice were anesthetized by intraperitoneal injection with a mixture of 0.75 mg/kg of medetomidine, 4.0 mg/kg of midazolam, and 5.0 mg/kg of butorphanol. Lung samples were dissected from mice and immediately immersed in RNA later (Invitrogen) stabilization reagent and stored at -80℃ for 24 hr. Samples were transferred to a new tube containing 1-mm glass beads and 700μL Trizol (Invitrogen). Tissues were subsequently homogenized using a MINI BEAD-BEATER (Biospec). Total RNA was extracted and purified using a RNeasy mini kit (QIAGEN, Tokyo, Japan) and quantified using a spectrophotometer (NanoDrop one, Thermo Fisher Scientific).  Total RNA was extracted from the lung as described above and checked for quality using RNA integrity scores were typically 7.0 and greater with an Agilent 2100 bioanalyzer (Agilent, Santa Clara, USA). For the quality control, mRNA had to meet the requirement of 28S/18S > 1, while miRNA had to meet the requirement of 28S/18S > 1.5. According to the manufacturer's protocol, the RNA was fragmented into small pieces. Afterward, the fragmented RNA was reverse-transcribed into cDNA and amplified with polymerase chain reaction (PCR) to create a cDNA library. Quality control and quantification of the libraries were performed with the Agilent 2100 bioanalyzer and real-time quantitative PCR (qPCR) (TaqMan Probe).
ENA first public
2024-10-01
INSDC center name
Department of rheumatology Tokyo Medical and Dental University (TMDU)
INSDC status
public
Submitter Id
E-MTAB-13838:lpr_Lung_Early3
age
19
broker name
ArrayExpress
collection date
not collected
common name
house mouse
developmental stage
adult
genotype
Lupus prone mice
geographic location (country and/or sea)
not collected
infect
Mouse-adapted SARS-CoV-2, 3 days post infection
isolate
not applicable
organism part
lung
sample name
E-MTAB-13838:lpr_Lung_Early3
scientific_name
Mus musculus
sex
female
strain
MRL/MpJ-lpr/lpr