Transcriptomic analyis follicular and marginal zone B ells after in vivo Notch2 blockade.

Marginal zone B cells which make rapid mitosis-independent plasma cell responses are dependent on Notch2 signaling for their generation and maintenance. In order to determine the effect of ongoing Notch signaling on the maintenance of the MZ transcriptome, we performed RNA-seq on MZ and follicular B cells after 12, 24 and 48 hours of in vivo Notch2 antibody blockade Overall design: Samples are sorted marginal zone and follicular B cells from animals after 12, 24 and 48 hours in vivo anti-Notch2 antibody treatment or isotype control. cDNA was prepared from RNA with 3' polyA capture method. Transcript abundance was measured by Kallisto psuedo-alignment and normalized (voom) and differential expression was assessed using limma.