Cxcr4f f

C57BL/6N mice were obtained from Nihon SLC (Hamamatsu, Japan). As previously described (Tanegashima et al., 2010 ), Cxcl14^−/− male and Cxcl14^+/− female mice with a C57BL/6 N background were crossed to produce Cxcl14^+/− and Cxcl14^−/− mice. LysM-Cre knock-in mice (Clausen et al., 1999 (link)) were obtained from RIKEN (RBRC02302), and CD11c-Cre (stock number 007567) and Cxcr4^F/F (stock number 008767) mice were obtained from the Jackson Laboratory (Bar Harbor, ME). Tlr9-KO mice (Hemmi et al., 2000 (link)) were obtained from Oriental Bioservice, Inc. (Kyoto, Japan).
BMDCs were prepared from 6-week-old C57BL/6N, Tlr9-KO, and Cxcr4-CKO mice by culturing bone marrow cells in RPMI-1640 medium (Nacalai Tesque, Kyoto, Japan) containing 10% fetal bovine serum (Thermo Fisher Scientific, Waltham, MA), mouse granulocyte-macrophage colony stimulating factor (10 ng/ml; Peprotech, Rocky Hill, NJ), and mouse IL-4 (10 ng/ml; Peprotech) for 10 days. Splenocytes were prepared from spleen removed from 6 to 8 week-old C57BL/6N mice. Red blood cells were lysed in RBC lysis buffer (140 mM NH₄Cl, 17 mM Tris-HCl, pH 7.6). All mice were housed in a pathogen-free animal facility under a 12 h light/dark cycle. All experimental procedures were pre-approved by the ethical committee of Tokyo Metropolitan Institute of Medical Science.

Wild-type, p21^−/−, Cxcr4^f/f, mTmG reporter, K14-Cre, K14-CreER, Tie-2Cre, Col2CreER, Rosa26-CreER, and J:NU mouse strains were obtained from Jackson Laboratory (strains 101045, 3263, 8767, 7676, 4782, 5107, 4128, 6774, 8463, and 7850, respectively). Mice were verified with genotyping instructions provided by Jackson Laboratory. For the ear wounding, we used a standard 2-mm mechanical punch to create a hole in the center of each outer ear (pinna) (Roboz). To delete reporter alleles, we administered 1 mg of tamoxifen (diluted in corn oil) daily by intraperitoneal injection for 5–10 d, depending on the Cre line. For Cxcr4^f/f mice, tamoxifen was administered for 5 d and then every other day for 1 mo. Mice used in this study were female age-matched littermates. All mice were housed in the animal facility of Stanford University on a 12-h light/dark cycle with ad libitum access to water and normal chow.