Fig. 6.

Potential metabolic regulators downstream of FR.A, site-specific gene set enrichment analysis (ssGSEA) was performed to interrogate the PTM signature databases (PTMsigDB) for both kinases and perturbations and plotted as nominal enrichment score (ES) versus -Log(adjusted p-value) for each cell line. CDK and Erk kinase activities were inversely correlated with FR treatment. The mTOR inhibitors rapamycin and torin1 were positively correlated with FR treatment. This analysis also identified a significant correlation with the EGFR inhibitor gefitinib, the MEK inhibitor, U0126, and the cMet inhibitor, SU11274 in MP46 cells. B, Torin-1 is an ATP-competitive mTORC1/mTORC2 dual inhibitor. Responses of MP41 and MP46 cells to torin1 were compared to FR in Agilent Seahorse assays for glycolytic stress. C, dosage responses to torin1 were measured in glycolysis and glycolytic capacity. MP41 cells showed no significant change in glycolysis but significant change in spare capacity (glycolytic capacity – glycolysis). D, responses of MP41 and MP46 cells to torin1 were likewise compared to FR in Agilent Seahorse assays for mitochondrial respiration stress. E, dosage responses to torin1 were measured in basal and maximal respiration and demonstrated highly similar responses in both cell lines. CDK, cyclin-dependent kinase; PTM, posttranslational modification.