To avoid the controversy data like GL261, we investigated the expression of TLR9 in U87 and U251 cells at both the mRNA level(RT-PCR) and protein level(immunofluoresence), our result for the first time showed that TLR9 staining located intracellularly and could be seen throughout the cytoplasm in U87 and U251 glioma cells. The gene discussed is TLR9; the disease is central nervous system cancer.