Figure 6

Inhibition of P2X7R with BBG reduced the number of glioma cells and microglia in human glioma cultures. Representative human glioma cell cultures before (A. control) and after (B. BBG) treatment with brilliant blue G. Red = isolectin GS IB4. Blue = DAPI nuclear stain. Scale bar, 20 μm. C. Treatment of human glioma cultures with 7 μM BBG at day 3 and day 5 post culture reduced the number of microglial cells. The number of microglia were counted using immunohistochemistry against a microglial marker, isolectin GS IB4 at 10 days after culturing. Nuclei were counter labeled with DAPI nuclear stain. Cell numbers were counted using MetaMorph imaging software. n = 109 randomly selected fields from 3 human gliomas that were not treated with BBG (control). n = 107 randomly selected fields from 3 human gliomas that were treated with BBG (BBG treated). D. Treatment of human glioma cultures with 7 μM BBG at day 3 and day 5 post culture reduced the number gliomas cells. The number of glioma cells were counted using immunohistochemistry against GFAP at 10 days after culturing. Nuclei were counter labeled with DAPI nuclear stain. Cell numbers were counted using MetaMorph imaging software. n = 114 randomly selected fields from 7 human gliomas that were not treated with BBG (control). n = 113 randomly selected fields from 7 human gliomas that were treated with BBG (BBG treated).