Additional file 6: of The regulation of combined treatment-induced cell death with recombinant TRAIL and bortezomib through TRAIL signaling in TRAIL-resistant cells

Figure S6. Combined treatment-induced cell death was not inhibited by the addition of autophagy inhibitors. (A, B) Cultured B16F10 cells were treated with ILz:rhTRAIL (100 ng/ml) and bortezomib (100 nM) with or without z-VAD-fmk, pan-caspase inhibitor: ILz:T, ILz:rhTRAIL; B, bortezomib; Z, z-VAD-fmk. After the indicated period, cells were harvested and cell lysates were prepared for immunoblotting analysis using appropriate antibodies: LC-3, microtubule-associated protein 1A/1B-light chain 3; Atg5, autophagy protein 5; p-S6, phosphorylated ribosome protein S6; p-S6K, phosphorylated ribosomal protein S6 kinase. (C) B16F10 and CT26 cells were cultured onto a 96-well plate and treated with ILz:rhTRAIL (100 ng/ml) and/or bortezomib (100 nM) following pre-treatment with Bafilomycin A1 (BfA1) for 3 h. Cell death was analyzed by XTT assay 24 h after the ILz:rhTRAIL and/or bortezomib treatment. (D) After pre-treatment with rapamycin (100 nM) or torin-1 (1 μM) for 3 h, ILz:rhTRAIL (100 ng/ml) and bortezomib (100 nM) were treated into cells: rapa, rapamycin. Cell death was analyzed by XTT assay 24 h after the ILz:rhTRAIL and bortezomib treatment. *p < 0.05 by Student’s t-test. (TIFF 236 kb