Figure 2.

Rines KO strategy and histology of Rines KO brain. A, Structures of the Rines protein, gene, targeting vector, and mutated allele. The sixth exon, which contains the RING finger domain (black box), was replaced with the neomycin resistance gene cassette (NEO), excised by Cre recombinase in germline cells and flanked by loxP sequences. The location of the 3′ and neoprobes used for Southern blotting and the expected size of the hybridizing fragments with 3′ and neoprobes after BglII digestion are shown. B, C, Confirmation of the mutant alleles by Southern blotting (B) and RT-PCR analysis (C). The locations of the 1-4 primers used for the RT-PCR analysis are shown. D–F, Survival rates, body growth, and brain histology of Rines KO mice. D, Genotype percentages in the 2162 3-week-old progenies from interheterozygote mating. There were no significant differences in the percentages of each genotype when we analyzed the data by a χ² test (p = 0.74). E, Body weight curves of male and female WT and Rines KO mice from weaning period (3 weeks) to 8 weeks. Male: WT, n = 19; KO, n = 21, Female: WT, n = 18; KO, n = 21. F, Cresyl violet staining of brain sections of 2-month-old WT and Rines KO mice. Scale bar, 1 mm. G, Dorsal views of WT and Rines KO mice at 7.5 months.