Figure 5.

G47Δ infection counteracts hypoxia-induced enrichment of CD133+ glioblastoma stem-like cells. (A): After 48-hour hypoxic culture, GBM4 (left panels) and GBM8 (right panels) cells were mock-infected (upper panels) or oncolytic herpes simplex virus (oHSV)-infected (enhanced green fluorescent protein-expressing G47ΔBAC; MOI = 0.2; lower panels) and cultured for 72 hours in hypoxia. CD133, GFP (infection), and 7AAD (cell death) were quantified by flow cytometry, and the values in each quadrant are the percentages of cells. (B): Western blot showing G47Δ-mediated abrogation of hypoxia-induced upregulation of CD133 protein in GBM4 (left panel) and GBM8 (right panel) cells. ICP4 (HSV immediate-early protein) bands demonstrate oHSV infection, and vinculin served as loading control. −, mock cell culture conditions (24 hours); +, G47Δ cell culture conditions (MOI = 1.5 for 24 hours). Abbreviations: 7AAD, 7-aminoactinomycin D; GBM, glioblastoma; GFP, green fluorescent protein; H, hypoxia; MOI, multiplicity of infection; N, normoxia.