Fig. 5. GBM-NS up-regulate CSPG4 in vivo.

(A) Xenograft gliomas originating from the GBM-NS CSPG4^ML (49%, BT168-NS) were explanted from engrafted mice and depleted of contaminant murine cells. Representative in vitro image shows GBM-NS 12 hours after isolation and murine cell depletion (upper panel) (scale bar, 150 μm). CSPG4 expression was assessed by flow cytometry (white histogram) compared to isotype control (black histogram). (B) BT168-NS were selected to obtain CSPG4^L and CSPG4^H cells using immunomagnetic sorting. Black histograms indicate the expression of CSPG4 in CSPG4^L and CSPG4^H cells as compared to unselected BT168-NS (white histograms). Data are representative of three different experiments. (C) Expression of CSPG4 (black histograms) in BT168-NS CSPG4^L and BT168-NS CSPG4^H versus unselected BT168-NS (white histograms) after 3 weeks in culture. (D) Gliomas originating from BT168-NS CSPG4^L and CSPG4^H cells were removed from engrafted mice. Upon depletion of contaminant murine cells, the expression of CSPG4 was measured by flow cytometry. Black histograms indicate the expression of CSPG4 in BT168 gliomas from CSPG4^L and CSPG4^H cells, whereas white histograms represent CSPG4 expression in CSPG4^L and CSPG4^H cells before in vivo implantation. (E) Kinetics of in vitro growth of CSPG4^L and CSPG4^H as measured by WST assay. (F) Kaplan-Meier survival curves of nude mice implanted intracranially with either BT168-NS CSPG4^L or BT168-NS CSPG4^H.