Transcriptome analysis of Regnase-1 deficient B cells at steady-state or upon B cell receptor stimulation

This study aims to identify the transcriptional programs that are regulated by the RNA binding protein Regnase-1 in B cells. To address the B cell specific roles of Regnase-1, we performed RNA seq analysis of B cells isolated from the spleens of mice in which Regnase-1 was deleted in a B cell specific manner using an inducible cre recombinase model, referred to as Regnase-1f/f hCD20TamCre, and compared to the appropriate controls, referred to as Regnase-1+/+ hCD20TamCre. Determining the differentially expressed genes from Regnase-1 deficient B cells compared to the control cells enabled identification of the transcriptional program regulated by Regnase-1 in B cells during steady state. In addition, we analyzed Regnase-1 deficient and control B cells that were stimulated with a stimulatory anti-IgM F(ab’)2 antibody that engages the B cell receptor (BCR). BCR engagement results into activation of B cells. Therefore, the differentially expressed genes from this analysis enabled identification of the genes that are modulated by Regnase-1 in B cells during an activated state. Transcriptome analysis was performed on 13 RNA samples which comprised 4 control samples from naïve B cells (Regnase-1+/+ hCD20TamCre), 3 samples from Regnase-1 deficient naïve B cells (Regnase-1f/fhCD20TamCre), 3 samples BCR stimulated control B cells (BCR-Regnase-1+/+ hCD20TamCre), and 3 samples from Regnase-1 deficient BCR stimulated B cells (BCR-Regnase-1f/fhCD20TamCre). BCR stimulation was done for 4 hours.