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. Author manuscript; available in PMC: 2015 Apr 1.
Published in final edited form as: Cancer Discov. 2014 Aug 6;4(10):1198–1213. doi: 10.1158/2159-8290.CD-14-0157

Figure 3. MPG loss sensitizes pGBM cells to alkylating agents.

Figure 3

a. Immunoblotting confirming MPG protein knockdown in pediatric GBM cell lines SJG2 and KNS42 following transfection with pooled stable clones generated from two unique MPG shRNA constructs.

b. Cell count of pediatric GBM cell lines SJG2 and KNS42 expressing MPG shRNA or shRNA controls (con) exposed to increasing doses of TMZ (0-500 uM). Cell counts were performed 72h post TMZ treatment.

c. Immunofluorescence of gamma H2AX in SJG2 cells expressing MPG shRNA or control (con) shRNA. Scale bar =16um.

d. Activated Cleaved caspase 3/7 assay of SJG2 and KNS42 cells treated with TMZ post 24h treatment. **p<0.01,***p<0.001

e. Colony forming unit (CFU) assay in SJG2 and KNS42 cells cultured with or without 100 uM TMZ. CFUs were counted after 14 days. **p<0.01 knockdown versus control cells.

f. Immunoblotting of MPG and LIG4 following siRNA treatment showing effective protein knockdown.

g-h. Plot of cell viability of SJG2 (g) and KNS42 (h) cells transfected with MPG, LIG4 or dual siRNA following exposure to varying concentrations of TMZ. Viability was measured using the almarBlue viability assay and quantified after 7 days. Dual siRNA knockdowns were also compared to single knockdowns to evaluate additive effects of double knockdown, *p<0.05,**p<0.01,***p<0.001.