162 | Chapter 6 Supplemental Figure 1. (A) Depiction of the gating strategy used to determine the amount of IgM+ cells after they have undergone successful V(D)J rearrangement. Primary WT bone marrow cells were cultured for 7 days on OP9 feeder cells, Flt3L, and IL7. Under these conditions on day 7, the cells acquire pro-to-pre B-cell phenotype assessed by FACS staining with B220-APC, CD34-PE, and IgM-PE/Cy7. On average 0.5% of the pre-B cells at this stage are IgM+ (B), 72 hours following the IL7 and Flt3L withdrawal, the portion of IgM+ cells increases to an average of 8%.
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