Figure S3. Generation and Characterization of the VH1-2/LC Mouse Model, Related toFigure 3
(A) Southern analysis of wild-type ES cells (lane 1) and VH1-2/LC ES clone (lane 2). The result verifiesJHdeletion (DJHb); additional restriction digest, not shown
here, indicated that the deletion occurred on theIgHballele.
(B) Southern analysis of wild-type ES cells (lane 1) and VH1-2/LC ES clone (lane 2). The result verifies the integration of VRC01LC intoJklocus (VRC01LC KI). The
VRC01LC, shown below the diagram, is composed of germline IGKV3-2001 (sequence in orange) and Jk1 from mature VRC01 (sequences in blue); two residues
in CDR L3, shown in black, cannot be assigned to either IGKV3-2001 or Jk1.
(C) HTGTS-rep-seq analysis of Vkusage in 129/Sv control and VH1-2/LC mouse B cells. The analysis was carried out as inFigures 1C and 1F. As mouseJk1-5on
oneIgkallele were deleted during the integration of VRC01LC, a primer downstream of Jk5 was used for the amplification of VRC01LC as well as Vk-Jk5 re-
arrangements on the intact mouseIgkallele. Given that the human IGKV3-2001 was inserted at the Jkregion, the slash sign on the x axis indicates the genetic
separation of IGKV3-2001 from the mouse Vksegments.
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