Figure 4.

CCL19 and CCL21 increase the migration of BMMs and osteoclasts. (a) BMMs were loaded to the upper chambers of transwell plates. CCL19 or CCL21 were added to the medium in the lower chambers. After 16 h, cells were fixed and stained with crystal violet. The migrated cells were counted. (b) BMMs were plated in Oris migration assay plates. The cells were incubated with M-CSF (30 ng ml⁻¹) and RANKL (100 ng ml⁻¹). After 2 days, the stoppers were removed, and the cells were washed twice with fresh medium. The cells were incubated with or without CCL19 (10 ng ml⁻¹) or CCL21 (10 ng ml⁻¹). After 16 h, the cells were fixed and stained with crystal violet. The stopper areas occupied with cells were quantified. (c) BMMs were transfected with control or CCR7 siRNA using Hiperfect. The levels of CCR7 mRNA and protein were assessed by PCR and western blots, respectively. (d) BMMs transfected with either CCR7 or control siRNA were loaded into the upper chambers of a transwell plate. CCL19 or CCL21 was added to the lower chambers. After 16 h, the cells were fixed and stained with crystal violet. Migrated BMMs were counted. BMM, bone marrow-derived macrophages; siRNA, small interference RNA.