Fig 1: Generation of Delta-24-GREAT. (A) Schematic representation of Delta-24-GREAT genomic structure. pCMV, cytomegalovirus promoter; BGH pA, bovine growth factor polyadenylation signal; ITR, inverted terminal repeat. (B and D) Expression in vitro of the GITR ligand (GITRL) on the surface of Delta-24-GREAT-infected murine glioma cells GL261-5 (100 MOIs) (B) and human brain tumor stem cells GSC17 (25 MOIs) (D) assessed by FACS, 48 hours after infection. Mock-infected cells stained with IgG isotype as primary antibody, and UV-inactivated Delta-24-GREAT-infected cells were used as control. Non-viable cells were excluded from the analysis using ethidium homodimer. Data are shown as mean ± SD of three independent experiments. α, antibody. (C and E) Expression of viral proteins, E1A and fiber, and murine GITRL, assessed by western blotting, in Delta-24-GREAT-infected murine glioma GL261-5 cells (C) and human glioma U-87MG cells (E) 48 hours after infection at the indicated MOIs. Mock, Delta-24-RGD, and UV-inactivated Delta-24-GREAT were used as controls at the highest MOI. GAPDH expression was used as loading control.