Figure 6.
The effect of in vitro melanoma cells-conditioned medium (CM) induced invasion of BMDCs in the AM signaling blockade. (A–C) In vitro regulation of BMDCs by A375, SK-MEL-28, and MeWo cells-CM. For all chambers, the bottom well was filled with melanoma cells-CM, while the control well was filled with DMEM containing 2% FBS (control). Immunoreactive AM secreted in CM was neutralized with pretreatment with αAM (70 μg/mL) for 30 min. αAMRs (70 μg/mL) or control IgG (70 μg/mL) was used to pre-treat bone marrow cells (5 × 105 cells), which were then placed in the upper chamber and incubated (see Materials and Methods). Migrated cells were stained with DAPI and counted under the microscope at 50× magnification. Numbers represent the number of migrated cells in 10 high-power fields, given as means ± SD of four independent experiments, each performed in triplicate. The asterisk (*) is used for comparison to control cells (** p < 0.01; *** p < 0.001) and the plus symbol (+) is used for comparison to CM-treated cells (+++ p < 0.001).