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The combined effects of MTX plus HDACIs were evaluated using a cell viability assay, mass spectroscopy imaging, and subcutaneous and intracranial xenograft models. HDACIs upregulated the ratio of FPGS/GGH expression resulting in increased polyglutamylation of MTX, but also downregulated expression of the target molecule of MTX DHFR. The combination of MTX and vorinostat decreased cell viability in vitro ( .05) and tumor volumes in a subcutaneous model ( .0001), and prolonged survival in an intracranial model ( .01), rel