Selection of H-CDR3 in the B cell development from SLC-knockout and C57BL/6 mice
Bone marrow and spleen cells were collected from pools of six mice enriched for B cells using CD19 MACS beads (Milltenyi Biotech), and sorted for pro-B (B220+c-kit+), pre-B (B220+c-kit-CD25+) immature (B220+CD93+IgM+) and FO (B220+CD93-43-21int23+) B cells. RNA was isolated using RNeasy kit (QIAGEN) and cDNA synthesis was performed according to the SuperScript II RT protocol (Invitrogen). Ig μ transcripts were amplified independently by PCR.