Figure 1.

Generation of GASP1 KO mice. (a) Targeting vector design for generating GASP1 KO mice. A cassette expressing the G418 resistance gene flanked by loxP sites was inserted into the intron upstream of the GASP1 open reading frame (ORF) (intron 4) and a third loxP site was inserted in the intron downstream of the GASP ORF (intron 5). ES cells from C57/Bl6 mice were transfected with this vector. Properly targeted clones (see b) were transfected with Cre-recombinase, and blastocysts from clones in which the GASP1 ORF was disrupted were implanted into C57/BL6 females. (b) Southern blotting analysis identified homologous recombination and single insertion using both (left) 5′ and (right) Neo probes. (c) Homogenates from wild-type (WT) and GASP1 knockout (KO) whole brain, cerebellum, spinal cord, and hypothalamus were analyzed by immunoblot (IB) and shows complete knockout of GASP1. Furthermore, there are no compensatory changes in the expression of the closest homolog, GASP2, in either of these regions.