Combined DDR kinase and mTORC1 inhibition bypasses AKT to induce BMF-mediated apoptosis. (A) A representative Eμ-Myc lymphoma (#4242) was transduced with a constitutively active AKT expression vector (Akt) or empty vector control (MSCV) and treated with increasing concentrations of BEZ235 for 24 hours prior to flow cytometric analysis for annexin-V/PI staining. (B) Lymphomas derived on a Bad- or Bim-deficient background and a Bad/Bim wild-type control (#4242) were treated for 24 hours with BEZ235 prior to analysis for annexin-V/PI positivity. (C) Schematic representation of the proposed kinase activities mediating BMF upregulation. Dashed lines indicate putative indirect interactions. (D) Lymphomas derived on a Bmf-deficient background (#14 and #31) and a Bmf wild-type control (#4242) were treated for 24 hours with BEZ235 prior to analysis for annexin-V/PI positivity. *P < .05 comparing genotypes (Bmf^+/+ vs Bmf^−/−; 2-way ANOVA) (E) Eμ-Myc lymphoma (#4242 MSCV/Bcl2) was treated for 24 hours with vehicle (DMSO), Everolimus (100 nM), DNA-PK inhibitor (KU57788; 1 μM), or combined Everolimus/KU57788 or BEZ235 (250 nM) prior to harvesting and preparation of protein lysates and immunoblotting for BMF. The fold-induction of BMF expression (densitometry relative to DMSO control and normalized to tubulin) is indicated above. (F) Lymphomas derived on a Bmf-deficient background (#14 and #31) and a Bmf wild-type control (#4242) were treated for 24 hours with Everolimus, KU57788, or combined Everolimus/KU57788 prior to analysis for annexin-V/PI positivity. # Indicates greater-than-additive effect for drug combination (SQ > 1). All results are representative of at least 3 independent experiments; bar graphs indicate mean (± SEM).