Cell - 8 September 2016

Figure S1. Generation and Characterization of the VH1-2 Mouse Model, Related toFigures 1and 3
(A and B) Results of Southern analysis designed to verify genetic modifications in VH1-2 mouse model. The diagrams, not drawn to scale, illustrate the restriction
digests and probes that were used to differentiate the region before (germline, GL) and after genetic modifications. (A) Southern analysis of liver DNA from VH1-2
mouse model (lanes 1-2) and wild-type 129/Sv mice (lanes 3-4). The result confirms homozygous replacement of VH81X with IGHV1-2*02 (Vh1-2) in VH1-2 mice.
(B) Southern analysis of liver DNA from VH1-2 mouse model (lanes 1-2) and wild-type 129/Sv mouse (lanes 3-4). The result validates IGCRI deletion (DIGCRI) on
both IgH alleles in VRC01HC mice.
(C) FACS analysis of splenic B cells from wild-type 129/Sv, VH1-2 and VH1-2/LC mice. In the left column, splenocytes were stained with anti-B220 and anti-Thy1.2
antibodies. Cells in the lymphocyte gate, as defined by FSC and SSC, were displayed in the FACS plot. The percentage of B cells (B220+) and T cells (Thy1.2+) are
indicated on the plot. In the middle column, splenocytes were stained with anti-B220, anti-IgM, and anti-IgD antibodies. B220+B cells were gated and shown in
the plot. The percentage of mature B cells (IgM+IgDhi) is indicated on the plot. In the right column, splenocytes were stained with anti-B220, anti-Igl, and anti-Igk
antibodies. B220+B cells were gated and shown in the plot. The percentage of Igk+and Igl+B cells is indicated on the plot. The data shown in this figure are
representative of at least three experiments for each genotype, as summarized in the table below.