Illumina HiSeq 2500 sequencing; RNASeq: Aryl hydrocarbon receptor governs a transcriptional programme that determines regulatory B cell differentiation and function
RNASeq: Aryl hydrocarbon receptor governs a transcriptional programme that determines regulatory B cell differentiation and function
Splenic CD21hiCD24hi B cells were isolated from Mb1cre and Ahrfl/-Mb1cre mice in the remission phase of arthritis. Sorted cells were either left untouched (ex-vivo) or stimulated with LPS+anti-IgM for 6h in IMDM media. Dead cells were excluded using DAPI.  Total RNA was isolated from these populations using the Picopure RNA isolation kit (ThermoFisher Scientific), according to manufacturer's instructions. 130-500ng of total RNA was fragmented followed by reverse transcription and second strand cDNA synthesis.  The double strand cDNA was subjected to end repair, A base addition, adapter ligation and PCR amplification to create libraries. Libraries were evaluated by Qubit and TapeStation. Sequencing libraries were constructed with barcodes to allow multiplexing of samples in 3 lanes.
Experimental Factor: stimulus
lipopolysaccharide and anti-IgM
Experimental Factor: phenotype
wild type