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Additional Supporting Information can be found in the online version of this article.

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ANA_22390_sm_suppinfofigure1.tif1819KSupplementary Figure 1: WST-1 proliferation data of stable HEK cells (A) and U138 cells (B) expressing GFP, IDH1wt-GFP or IDH1R132H-GFP. Error bars represent the Standard Deviation of the Mean. * = p<0.05, ** = p<0.01
ANA_22390_sm_suppinfofigure2.tif1692KSupplementary Figure 2: Light emitted by tumors of mice injected with GFP expressing U87 cells (n=10, green), IDH1wt-GFP expressing U87 cells (n=10, blue) and IDH1R132H-GFP expressing U87 cells (n=10, red) as measured by IVIS.
ANA_22390_sm_suppinfofigure3.tif2515KSupplementary Figure 3: Whole brain slices (upper lane), H&E stained slices (middle lane) and GFP stained slices (lower lane) of mice injected with U87 cells expressing IDH1 wt-GFP (left) or IDH1 R132H-GFP (right).
ANA_22390_sm_suppinfofigure4.tif4320KSupplementary Figure 4: NADPH producing enzymatic activity of U87 cells overexpressing IDH1R132H-GFP (red line) or IDH1wt-GFP (blue line) after adding different concentrations of isocitrate (A: 4 μM; B: 8 μM; C: 40 μM). Datapoints are normalized on actin expression (see Supp Fig 5A). D. NADPH consuming enzyme activity of U87 cells expressing IDH1R132H-GFP (red line) or IDH1wt-GFP (blue line) after adding a-ketoglutarate. Datapoints are normalized on actin expression (see Supp Fig 5B).
ANA_22390_sm_suppinfofigure5.tif1756KSupplementary Figure 5: Western blots measuring actin expression in U87 cell pellets used for normalization of the enzyme measurements in Supp Fig 4A to C (A) and 4D (B). C. Western blots showing protein levels of the isolated IDH1wt-GFP and IDH1R132H-GFP enzymes, the co-transfection of both enzymes and GFP, as used for normalization in Fig 5. Rat IDHC is used as a control.
ANA_22390_sm_suppinfofigure6.tif2157KSupplementary Figure 6: NADPH producing enzymatic activity of patient-derived IDH1R132H mutant tumors (red line; n=5) and IDH1wt tumors (blue line; n=5) after adding different concentrations of isocitrate (A: 4.4 μM; B: 8.8 μM; C: 44 μM). Datapoints are normalized on by protein concentration as measured by a Bradford protein assay.
ANA_22390_sm_suppinfofigure7.tif2934KSupplementary Figure 7: A. D-2-HG concentration measured in the medium of U87 cells expressing GFP, IDH1wt-GFP or IDH1R132H-GFP. B. D-2-HG per mg protein measured in cell pellets of U87 cells expressing GFP, IDH1wt-GFP or IDH1R132H-GFP. C. D-2-HG concentration measured in tumors (n=5 per group) of mice injected with U87 cells expressing GFP, IDH1wt-GFP or IDH1R132H-GFP.
ANA_22390_sm_suppinfofigure8.tif2725KSupplementary Figure 8: Network constructed by Ingenuity of all seven genes (in grey) that are differentially expressed by cell lines (U87 and U138) expressing IDH1R132H-GFP compared to cell lines expressing IDH1wt-GFP.
ANA_22390_sm_suppinfofigure9.tif1783KSupplementary Figure 9: p-Akt expression in U87 cell lines overexpressing GFP, IDH1wt-GFP or IDH1R132H-GFP.
ANA_22390_sm_suppinfo.doc43KSupporting Information

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