organism: Mus musculus
description: Protocols: Neonatal mouse brains were dissected out, dissociated into single cells, and cultured in DMEM with 10 ng/mL GM-CSF, 10% FBS, 50 U penicillin, and 50 mg/mL streptomycin at 37°C. Collected primary microglia were treated with 100 ng/ml LPS for 3 hours. Microglia were transfected with either scrambled control siRNA or Igf2bp1 siRNA for 48 hours afterwards. The genome-wide m6A was detected by RIP kit (Millipore). RIP Lysis Buffer was added and the products were lysed on ice and collected. Then, the beads were washed with RIP Wash Buffer. After removal of the supernatant, the beads were incubated with 1 μg antibodies of m6A (56593S, CST),for 30 min at room temperature, and normal rabbit immunoglobulin G (Abcam) was used as the control. Subsequently, the cell lysate was incubated with RIP buffer containing magnetic beads. RNA was extracted after proteinase K digestion. Five μg RNA of samples were used for library preparation by NEBNext Ultra Directional RNA Library Prep Kit for Illumina (NEB, Ipswich, MA, USA) following manufacturer's instructions.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2022-12-31
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2022-12-31
attributes.tag: External Id
attributes.value: SAMEA12814114
attributes.tag: INSDC center alias
attributes.value: TONGJI UNIVERSITY
attributes.tag: INSDC center name
attributes.value: TONGJI UNIVERSITY
attributes.tag: INSDC first public
attributes.value: 2022-12-31T00:57:48Z
attributes.tag: INSDC last update
attributes.value: 2022-12-31T00:57:48Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: RNA interference
attributes.value: Igf2bp1 targetting siRNA
attributes.tag: Submitter Id
attributes.value: E-MTAB-11411:LPS+Igf2bp1 siRNA input 2 R1
attributes.tag: age
attributes.value: 1
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell_type
attributes.value: microglial cell
attributes.tag: common name
attributes.value: house mouse
attributes.tag: developmental stage
attributes.value: mouse postnatal
attributes.tag: disease
attributes.value: neuroinflammation
attributes.tag: genotype
attributes.value: wild type genotype
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: organism part
attributes.value: brain
attributes.tag: sample name
attributes.value: E-MTAB-11411:LPS+Igf2bp1 siRNA input 2 R1
attributes.tag: strain
attributes.value: C57BL/6J