Fig. 5

IFNγ and Smac mimetics synergistically induce apoptosis in HCC827 NSCLC cell line. a, b HCC827 NSCLC cells were incubated with 1–25 units of human IFNα or 1–25 ng/ml of IFNγ, IFNλ or TNFα (a), or with TRAIL (200 or 400 ng/ml) (b) in the presence or absence of 20 nM AZD5582 for 48 h. c HCC827 cells were incubated with various doses of poly(I:C) in the presence or absence of 20 nM AZD5582 for 48 h. d, e HCC827 cells were incubated with 5 ng/ml IFNγ plus different doses of SM164 or AZD5582 for 48 h. The remaining adherent cells after 48 h treatment with AZD5582 (20 nM) plus IFNγ (5 ng/ml) were subjected to Annexin V apoptosis assay (e). f HCC827 cells were treated with IFNγ (5 ng/ml) and AZD5582 (20 nM) for 7 or 26 h, and cell lysates at equal amounts were subjected to Western blotting with indicated antibodies. g, h HCC827 cells were treated with IFNγ (5 ng/ml) and AZD5582 (20 nM) for 24 h and caspase-8 and -3 activities were determined. i HCC827 cells were incubated with DMSO, AZD5582 (20 nM), IFNγ (5 ng/ml) or AZD5582 plus IFNγ (AZD + IFNγ) in the presence DMSO, Z-VAD-FMK (25 µM), necrostatin-1 (Nec-1, 25 µM), TNFα neutralizing antibody (TNFαAb, 1 µg/ml), GSK872 (GSK, 5 µM), necrosulfonamide (NSA, 1 µM), or VX-765 (10 μM) for 48 h. Cell viabilities (a–e, i) were assessed by MTS assay and cell survival rates were calculated by comparison to DMSO-treated control cells (n = 3). **p < 0.01; ***p < 0.001 versus DMSO. Results represent three independent experiments