Mice Lacking Expression of Secondary Lymphoid Organ Chemokine Have Defects in Lymphocyte Homing and Dendritic Cell Localization

Gunn, Michael D.; Kyuwa, Shigeru; Tam, Carmen; Kakiuchi, Terutaka; Matsuzawa, Akio; Williams, Lewis T.; Nakano, Hideki

Mice. BALB/c-plt / plt mice were produced by backcrossing plt mice 10 generations into a BALB/c genetic background as described previously (29) . BALB/c control mice were obtained from The Jackson Laboratory. All experiments were performed on 6-10-wk-old age-and sex-matched mice maintained under specific pathogen-free conditions. RNA Expression Studies. For Northern analysis, mRNA from tissues of wild-type and plt mice was subjected to gel electrophoresis, transferred to Hybond-N ϩ membranes (Amersham Pharmacia Biotech), and probed using randomly primed mouse SLC, ELC, and actin cDNA. For in situ hybridizations, paraffin sections (5 m) from ϩ / ϩ and plt mice were deparaffinized, fixed in 4% paraformaldehyde, and treated with proteinase K. After washing in 0.5 ϫ SSC, the sections were covered with hybridization solution, prehybridized for 1-3 h at 55 Њ C, and hybridized overnight with sense or antisense 35 S-labeled riboprobe transcribed from the mouse SLC or ELC cDNA. After hybridization, sections were washed at high stringency, dehydrated, dipped in photographic emulsion NTB 2 (Eastman Kodak Co.), stored at 4 Њ C for 8 wk, developed, and counterstained with hematoxylin and eosin.