All animal procedures were performed within the guidelines of the Animal Care and Experimentation Committee of Gunma and Osaka University, and all animals were bred at the Institute of Animal Experience Research of Gunma and Osaka University.
Generation of Rab8b knockout (BKO) mice was performed largely according to a previous report (Harada et al., 1994 (link)). Rab8b genomic clones were isolated from a mouse genomic BAC library from the 129Sv/J mouse strain (RPCI-22: Children's Hospital Oakland Research Institute, Oakland, CA), using a fragment of the mouse Rab8b second intron (indicated as probe 2 inFig. 1A ) as a probe. The targeting vector consisted of a 1-kb 5′ homologous region, a SA-IRES-βgeo-polyA cassette flanked with two FRT sites and a downstream loxP site (Sato et al., 2007 (link)), a 1-kb genomic sequence region including exon 2 (from 350 bp upstream of exon 2 to 400 bp downstream of exon 2), a single loxP site, and a 6.7-kb 3′ homologous region. Two targeted clones (B19, C21) were identified by Southern blot analysis using probes 1 and 2 (Fig. 1B,C ). To generate the null mice, we crossed Rab8b βgeo/+ mice with Act-Flp-e transgenic mice and then with CMV-cre transgenic mice (Jackson Laboratory, Bar Harbor, ME).
Generation of Rab8b knockout (BKO) mice was performed largely according to a previous report (Harada et al., 1994 (link)). Rab8b genomic clones were isolated from a mouse genomic BAC library from the 129Sv/J mouse strain (RPCI-22: Children's Hospital Oakland Research Institute, Oakland, CA), using a fragment of the mouse Rab8b second intron (indicated as probe 2 in