biostudies-arrayexpressMetabolomicsUnknownTranscriptomicsGenomicsProteomicsFrancesco StrozziNAIllumina NovaSeq XChromium 10X GenomicsRNA-seq of coding RNA from single cellsHomo sapiensHomo sapienshttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-15419Single cell RNAseq analysis of in vitro B-cells stimulated with CpG, CGS and Inupadenantbiostudies-arrayexpressSample Collection - Tonsil sample collected by adult patients undergoing tonsillectomy was cut into small fragments before mechanical dissociation through a 100 mm cell strainer using a syringe plunger. Tonsillar mononuclear cells (TMNC) were then isolated by lymphoprep density gradient centrifugation using SepMate 50 tubes (STEMCELL Technologies) and tonsillar B cells were enriched using EasySep™ Human Pan-B Cell Enrichment Kit (STEMCELL Technologies) following the manufacturer’s protocol. Isolated B cells were cultured in a 24-well plate at a concentration of 2 million per well in 1ml of X-vivo medium supplemented with 5% inactivated human serum, 1mM of Na-pyruvate and 1mg/ml of CpG for 3h at 37°C, 5% CO2. The A2AR agonist CGS-21680 was added at 5 mM alone or in combination with the A2AR antagonist inupadenant (EOS100850) at 300 nM final concentration.Library Construction - Library preparation was performed following the Chromium FixedRNAProfiling_MULTIPLEXED Samples (10X Genomics) protocol. Chromium Fixed RNA Kit, Human Transcriptome, 4 rxns x 16 BC, PN-1000476Sequencing - Sequencing was performed on Illumina NovaSeq, generating 2x150 paired-end readsNucleic Acid Extraction - Treated cells were then fixed 1h at room temperature following the Fixation of Cells & Nuclei for Chromium Fixed RNA Profiling (10X Genomics) manufacturer’s protocol.OrganizationMINSEQE ScoreAssays and DataMAGE-TAB FilesPaola TieppoDavid CarbonezFrancesco StrozzifalseSingle cell RNAseq analysis of in vitro B-cells stimulated with CpG, CGS and InupadenantSingle cell RNAseq analysis of in vitro B-cells stimulated with CpG, CGS and Inupadenant2025-12-07T00:00:00Z2025-12-07T02:01:58.36Z2025-07-30T15:33:02.987ZE-MTAB-15419ERP177492EFO_0002944EFO_0004170EFO_0005684EFO_0005518EFO_0004184