Figure 6

IgH recombination detected upon culture of tamoxifen treated E2A^ER/ER pre-proB cells. DNA was analyzed for IgH V to DJ rearrangements from Day 8 cultured wild-type (WT), Rag1^-/- (Rag) and E2A^ER/ER (ER) pre-proB cells using V_H1 and J_H4 primers. Cells were cultured in hormone-free media on S17 stromal cells with IL-7 and tamoxifen (T) or DMSO (D). V-DJ rearrangement products using V_H1 family gene segments and J_H4 are shown. WT and Rag were used as positive and negative controls, respectively. No products were detected when using dH₂O as an additional negative control (data not shown). CD14 was used to demonstrate DNA loading. This result is representative of 4 independent nested PCR reactions.