SUPPLEMENTAL EXPERIMENTAL PROCEDURES Mutant Mouse Strains

were generated by homologous recombination in embryonic stem cells. Sequences used for the generation of the CAPS-2 targeting vector were amplified by PCR with engineered primers from a pBeloBAC11 derived BAC clone (23D20) containing 182,820 bp of the 129/SVJ murine CAPS-2 gene, including exons 2-6. Genomic sequences were subcloned into pTK-Neolox (Figure S1A). In the targeting vector a 500 bp genomic fragment containing exon 5 (Cisternas et al., 2003) was replaced by a loxP-flanked Neomycin resistance gene. The vector also contained two copies of the herpes simplex virus thymidine kinase for negative selection. Recombinant stem cell clones were identified by Southern blotting (Figures S1A and S1B). One of a total of four positive clones was injected into mouse blastocytes to obtain highly chimaeric mice that transmitted the mutant gene through the germ line. Germ line transmission of the mutant gene was confirmed by Southern blotting (Figure S1B). Subsequent routine genotyping was performed by PCR. Western blot analyses of adult mouse brain homogenates with a CAPS-2 specific antibody (Speidel et al., 2003) demonstrated the total absence of CAPS-2 in homozygous mutant brains (Figure S1C). CAPS-1/CAPS-2 double KO