IKBKE/TBK1 inhibition modulates MYC signaling pathway. (A) GSEA of shIKBKE/TBK1-sensitive vs -resistant AML cell lines. Gene set databases used are c6 oncogenic signature and c1 hallmarks. (B-D) MOLM14 or OCIAML5 cells were treated with momelotinib (CYT; 2.5 µM), MRT (2.5 µM), BX795 (2.5 µM), or ruxolitinib (10 µM) for 6 hours, after which RNA or protein was isolated. 18S RNA was used as an internal control for RNA quantification. For OCIAML5, cells were cultured without GM-CSF for 24 hours before drug treatment. (E-G) MOLM14 or OCIAML5 cells were infected with lentiviral vectors expressing shRNA targeting IKBKE, TBK1, or luciferase (as nontargeting control). After 54 hours of puromycin selection, RNA or protein was isolated. For RT-PCR, 18S RNA was used as an internal control. Data are mean ± SE of 2 independent experiments. Quantitation of MYC:tubulin ratio (F) was performed using ImageJ software. RUXO, ruxolitinib.