Extended Data Fig. 1: Early B-cell development in Tcf3Aid/Aid, Ebf1Aid/Aid, Pax5Aid/Aid, Ikzf1Aid/Aid, and Ikzf3Aid/Aid mice.
a, The indicated C-terminal tag sequences, consisting of a glycine linker, a minimal auxin-inducible degron (Aid) sequence65 and the V5 epitope, were added in frame to the last amino acid of E2A, Pax5, Ikaros, or Aiolos. The stop codon is denoted by an asterisk. b, Schematic diagram describing the generation of the Ebf1Aid allele. The exon-intron structure of the 5′ region of the Ebf1 gene is shown together with the sequences encoding the V5 epitope, Aid sequence, glycine linker and the exon 1-encoded amino acid sequence inserted into the 5′ part of exon 2 of the Ebf1Aid allele (for DNA sequence see Supplementary Table 5). c, Analysis of early B-cell development in the bone marrow of Tcf3Aid/Aid, Ebf1Aid/Aid, Pax5Aid/Aid, Ikzf1Aid/Aid, Ikzf3Aid/Aid, and control (Ctrl) wild-type mice at the age of 5 weeks. The flow-cytometric analysis of the different mice was performed on the same day in one experiment. The percentage of cells is indicated next to the gate of the different B-cell types. One of three experiments is shown. Imm, immature; Rec, recirculating. d, Expression of the indicated Aid-tagged TFs in ALPs, BLPs, and pro-B cells, which was determined by flow-cytometric analysis of intracellular staining with an anti-V5 antibody. The histogram displays the protein levels of the different TFs in the three progenitors. The flow-cytometric definition of the different B-cell types (c,d) is also described in Methods.
