FIGURE 4.

Generation of AS PKCδ knock-in mice. A, organization of the mouse Prkcd gene, targeting constructs, and M425A mutant allele resulting from homologous recombination. Filled boxes, exons 7–18. Met-425 is in exon 13. Arrowheads, loxP sequences. The targeting vector was composed of a 3.1-kb short arm carrying the M425A mutation, a Neo expression cassette flanked by two loxP sites, a 5.4-kb-long arm, and a diphtheria toxin A (DTA) gene. B, verification of genotypes by PCR using the G8-PCR-F and G8–35-R primers. The wild type allele (+) generates a 0.47-kb product, and the M425A mutant allele (A) generates a 0.6-kb product. C, Western blot showing similar PKCδ immunoreactivity in neutrophils from WT and AS PKCδ knock-in (KI) mice.