Figure 6. Combined loss of MPG and ATM sensitizes pGBM cells to TMZ in vivo.

a. Immunoblotting of SJG2 cells demonstrating effective MPG, ATM or dual knockdown.

b. Kaplan Meir survival curve analysis of intracranial injected SJG2 cells expressing control shRNA, MPG, ATM or dual knockdown into NOD-SCID mice.

c. Kaplan Meir survival curve analysis of intracranial injected SJG2 cells expressing control shRNA, MPG, ATM or dual knockdown into NOD-SCID mice treated with TMZ (65 mg/kg/5 days).

d. Quantification of ki67 staining in mice (n=3 mice per group) from c.

e. Kaplan Meir survival curve analysis of intracranial injected SJG2 into NOD-SCID mice treated with vehicle, TMZ (65mg/kg/5days), Methoxyamine (MA 100mg/kg/5days) and both TMZ+MA. Mice were treated upon confirmation of tumour by T2-MRI.

f. Quantification of ki67 staining in mice (n=3 mice per group) from e.

g. Kaplan Meir survival curve analysis of an orthotopic patient derived xenograft (PDX) mouse model from pGBM 462 cells. 20 mice were injected with 5 per each treatment arm: Vehicle, TMZ (65mg/kg/2 weeks), Methoxyamine (MA 100mg/kg/2 weeks) and both TMZ+MA (65mg/kg TMZ and MA 100mg/kg for 2 weeks). Mice were treated upon confirmation of tumour by T2-MRI.

h. Hematoxylin and eosin stain (H&E stain) of represented tumors mice from each treatment arm confirming high grade glioma/GBM morphology. Scale bar, Large insets at 500um and small magnified insets at 25um. N denotes normal mouse brain and T denotes xenograft tumor growth.