Fig. 1

Expression of CIC and its targets in human GBM tumors and cells. Human operative GBM samples or normal derived brains (NB) were lysed and a immunoblotted with indicated antibodies b or total RNA extracted and quantitative real-time PCR analysis was carried out using TaqMan gene expression assays. The graph depicts fold changes in CIC expression relative to normal brain. c Nuclear or cytoplasmic lysates were isolated from human operative GBM samples or normal brain and were immunoblotted with indicated antibodies. d Human operative astrocytoma samples were lysed and immunoblotted with indicated antibodies. Human-derived GBM cell lines or normal human astrocytes (NHA) were lysed and e protein lysates were immunoblotted with indicated antibodies (f) or total RNA extracted and quantitative real-time PCR analysis was carried out using TaqMan gene expression assays. The graph depicts fold changes in CIC expression relative to NHA. Data represent mean ± s.e.m. of three independent experiments performed in triplicate. *P < 0.05 Student’s t-test compared with NHA. g GL261 cells or normal mouse astrocytes were lysed and protein lysates were immunoblotted with indicated antibodies. Glioma stem cells (GSCs) were lysed and h protein lysates were immunoblotted with indicated antibodies (i) or total RNA extracted and quantitative real-time PCR analysis was carried out using TaqMan gene expression assays. The graph depicts fold changes in CIC expression relative to normal neural stem cells (NSC). Data represent mean ± s.e.m. of three independent experiments performed in triplicate. *P < 0.05 Student’s t-test compared with NSC. Representative images of hematoxylin and eosin (H&E) staining and immunohistochemistry using anti-CIC antibody of sections obtained from brains of intracranial xenograft (j) (GSC (8-18)) scale bar, 1 mm, or k GL261 mice scale bar, 500 μm. l Tumors or unaffected normal brains obtained from intracranial U87 xenograft were lysed and protein lysates were immunoblotted with indicated antibodies. m High-grade tumors obtained from two different oncogenic HA-H-Ras(12 V) knock-in RasB8 transgenic mice or from two different normal tissue obtained from wild-type background mice were lysed and protein lysates were immunoblotted with indicated antibodies. The immunoblot data are representative of at least three separate experiments