RNA-seq analysis of bead-purified B cells of wild-type BALB/c mice stimulated in vitro with TLR9 agonists (CpG ODN 1826, Hokkaido System Science) or left unstimulated.

The goal of this study was to develop a B-cell specific gene signature to catalog induced genes after short term in vitro TLR9 stimulation of wild-type BALB/c B cells. B cells were bead purified from splenocytes (two spleens combined for one replicate) of 7-11-week-old wild-type BALB/c mice. The bead purified B cells (10 million (M) per replicate) were stimulated for 4 hours at 10M/ml with TLR9 agonist CpG (CpG ODN 1826, Hokkaido System Science) at 10µg/ml or left unstimulated. RNA was isolated using the RNeasy Plus Mini Kit (QIAGEN). Samples were sequenced on an NextSeq500 (Illumina, Inc) with 2 x 75 bp paired-end reads (20 million reads per sample). B-cell specific gene signature of induced genes after short term in vitro TLR9 stimulation was cataloged. Overall design: mRNA profiles of B cells of wild-type BALB/c mice stimulated in vitro with TLR9 agonists (CpG ODN 1826, Hokkaido System Science) or unstimulated, were generated in three replicates, by sequencing on an Illumina NextSeq500 using PE 150 Cycles High Output flowcell.