Anti-OPN antibodies for western blotting and immunohistochemical staining were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA) and Developmental Studies Hybridoma Bank (DSHB, IA, USA), respectively. Anti-Ki-67 and anti-VEGFR2 were from Cell Signaling Technology (MA, USA). Rabbit anti-vascular endothelial growth factor (VEGF) antibody was from Thermo Scientific (MA, USA). Mouse anti-rat endothelial cell antigen (RECA) antibody was from Abcam (MA, USA).
Rat C6 glioma cell line was purchased from the Bioresource Collection and Research Center (ATCC #CCL-107, BCRC, Taipei, Taiwan). The cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Gibco, NY, USA) supplemented with 10% FBS (Gibco) and 1% penicillin-streptomycin (Gibco), and maintained under tissue culture condition at 37 °C with 95% air, 5% CO2, and 100% humidity.
Rat C6 glioma cell line was purchased from the Bioresource Collection and Research Center (ATCC #CCL-107, BCRC, Taipei, Taiwan). The cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Gibco, NY, USA) supplemented with 10% FBS (Gibco) and 1% penicillin-streptomycin (Gibco), and maintained under tissue culture condition at 37 °C with 95% air, 5% CO2, and 100% humidity.