Figure 3.

Mast cells (MCs) can significantly induce proliferation and invasion of brain metastasis (BM) cells. The BM cell lines were cocultured with MCs or cultured alone for 24 and 48 h and (A) AlamarBlue cell proliferation assay performed with NCI-H1915 (left panel) and U3333MET (right panel) cultured alone (red) or in coculture with LAD2 cells (green). (B) EdU was added to the cultures 4 h before the experiment endpoint and the cells were processed for immunofluorescence to analyze incorporated EdU (left panel). The results are expressed as EdU positive cell numbers (right panel). (C) Migration capacity of the BM cells was assessed by wound healing assay after 24 and 48 h when grown alone or in coculture with MCs (left panel). The results are shown as percentage of healed wound area (right panel). The experiments were performed three times in triplicates and mean values + SEM was plotted, ns, not significant; **p < 0.01, ***p < 0.001.