title: IgG_LPS_N3_R1
organism: Mus musculus
description: Protocols: In vivo, platelet depletion was performed in female C57BL/6J mice (12 weeks) by intravenous (i.v.) injection of 2 mg/kg of polyclonal rat anti-mouse GPIbα (R300) or same amounts of a polyclonal rat IgG none-immune (C301) antibody as control (Emfret Analytics). Mice were then challenged 12 hours later with i.v. injection of 2 mg/kg LPS or PBS as control. Blood was collected after 3 h, and PBMCs were isolated by density gradient centrifugation in Ficoll Paque PLUS. Mouse monocytes were separated from PBMCs by FACS-sorting gating on Ly6G−, IA/IE−, CD45+, CD11b+, and CD115+ cells and sorted directly into QIAzol lysis reagent. Lysates were frozen in liquid nitrogen and sent to GENEWIZ Azenta Life Sciences for bulk RNA sequencing.   For bulk RNA sequencing, samples were processed according to GENEWIZ (Azenta Life Sciences) using TRIzol extraction protocol. QC was done on QuBit and TapeStation.   RNA extraction was performed, followed by a library preparation (Azenta, GeneWiz).  SmartSeq HT library preparation kit with maximum input volume as the RNA amount was below QuBit detection limit.  mRNA sequencing via polyA selection: 1) mRNA Enrichment, mRNA Fragmentation, and Randon Priming; 2) First and second strand cDNA synthesis; 3) End Repair, 5' Phosphorylation, and dA-Tailing; 4) Adaptor Ligation, PCR Enrihment, and Sequencing.
attributes.tag: ENA first public
attributes.value: 2024-06-15
attributes.tag: INSDC center name
attributes.value: Institute of Innate Immunity, Medical Faculty, University of Bonn
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-14126:IgG_LPS_N3_R1
attributes.tag: age
attributes.value: 12
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell_type
attributes.value: FACS-Sorted Ly6G-I-A/I-ECD45+ CD11b+CD115+ monocytes
attributes.tag: collection date
attributes.value: not collected
attributes.tag: common name
attributes.value: house mouse
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: genotype
attributes.value: wild type genotype
attributes.tag: geographic location (country and/or sea)
attributes.value: not collected
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: organism part
attributes.value: blood
attributes.tag: replicate
attributes.value: 5
attributes.tag: sample name
attributes.value: E-MTAB-14126:IgG_LPS_N3_R1
attributes.tag: scientific_name
attributes.value: Mus musculus
attributes.tag: sex
attributes.value: female
attributes.tag: stimulus
attributes.value: LPS
attributes.tag: strain
attributes.value: C57BL/6J
attributes.tag: treatment
attributes.value: IgG