AS602868 inhibits constitutive activation of IKK2 and NF-κB in AML blasts. Blasts from patients 8, 17, and 18 were incubated with indicated doses of AS602868 for 18 hours. Blasts from patients 4, 6, 11, 12, 15, and 16 were incubated with 10 μM AS602868 for 18 hours or left untreated. (A, left column) AS602868 blocked autophosphorylation of IKK2 in AML blasts. Cell lysates were resolved by SDS-PAGE and transferred to Immobilon membranes. The autophosphorylation status of IKK2 was analyzed with antibodies against phosphorylated serines 177 and 181, and anti-IKK2 antibodies show equal amounts of the kinase in the different conditions. AS602868 inhibits NF-κB activation (Panel A, right column; and panel B). Total cell extracts were incubated with radioactive probes encompassing the NF-κB or OCT-1 DNA binding sequences. Complexes were separated by nondenaturing electrophoresis followed by autoradiography (EMSA). ns indicates nonspecific.