Co-inhibition of NF-κB and JNK is synergistic in TNF-expressing human AML. (A) CFU from human LC lines treated with indicated doses of SP, BAY, or combination. (B) Nuclear fractionation analysis showing NF-κB activity in AML cell lines. (C) Several TNF-high–expressing AML cell lines were treated with TNF and BAY individually or in combination. Levels of p-JNK and p-cJUN were examined by Western blotting. (D) Primary AML cells were treated with indicated doses of BAY, SP, or combination, and then assessed for CFU. Right axis indicates relative TNF mRNA expression levels relative to average healthy donor PB stem cells (PBSCs; n = 3). mRNA transcripts were set to GAPDH as control. ∼ indicates increased TNF mRNA level compared with PB stem cell control, P < 0.05. * indicates P < 0.05 when compared with vehicle-treated control, unless otherwise noted, as determined by one-way ANOVA. # indicates P < 0.05 significant difference when compared with indicated conditions. Values shown are mean values ± 1 SD from three independent trials. B and C are representative results from three independent trials. (E) Proposed mechanism for TNF function in AML.