Generation of
Lsp1−/−mice. (A) Restriction maps of pTEX1 and of the exon 1-containing regions of the wt and mutantLsp1 alleles. Thick lines represent regions of homology between the vector and endogenous allele. Diagnostic BamHI and EcoRI fragments, which hybridize with probes A and B on genomic Southern blots, are indicated in kilobases. Neoindicates neomycin gene; tk, HSV-thymidine kinase gene; K,KpnI; E, EcoRI; C, ClaI; B,BamHI, X, XbaI; S, SstI (maps not drawn to scale). (B) Southern blot analysis of targeted ES cell DNA (lanes 1, 5, and 9) and of tail DNA from F2 littermates (lanes 2-4, 6-8, and 10-12). Hybridizing bands from the wt and mutant (mu) allele are indicated. Band x is a cross-hybridizing BamHI fragment that is also detected in wt ES cells (not shown). (C) Cell lysates (1 × 105 cell equivalents) from bone marrow (BM), thymus (Th), spleen (Sp), and peritoneal cavity (Per) of wt (odd numbered lanes) and Lsp1−/− (even numbered lanes) mice were analyzed using antimouse LSP1 serum or antiactin mAbs as described in “Materials and methods.” The positions of the markers in kilobases and kilodaltons are indicated on the right side of the panels in B and C, respectively.
