JL5 but not JL12 regulates BMP signaling and induces cell death. a, b Western blot analyses of H1299 cells treated with JL5 or JL12 for 72 h showing that JL5 but not JL12 decreases expression of Id1, XIAP, and pTAK1. c H1299 cells stably expressing the Id1-luciferase reporter were treated with JL5 and JL12 for 48 h. JL5 but not JL12 decreases the activity of the Id1 promoter. d, e H1299 cells were treated with JL5 or JL12 for 72 h and (d) the percent dead and (e) the number of live cells determined. f, g H1299 cells were treated with JL5 for 7 days and the percent dead and number of live cells determined. h Western blot analysis of H1299 cells treated with JL5 for 3 days demonstrating that JL5 activates caspase-3 and cleaves PARP. i H1299 cells were treated with JL5 2.5 μM for 48 h. Cells were stained were then stained with fluorescently labeled Annexin V and 7-AAD and analyzed by flow cytometry. JL5 induced both late stage apoptosis and necrotic cell death. j TUNEL assay of H1299 cells treated with JL5 for 24 h demonstrating an increase in DNA double-stranded breaks. k H1299 cells were pretreated with Z-VAD-FMK (VAD) or necrostatin for 1 h then treated with DMSO or JL5 for 3 days and percent death cells determined. VAD but not necrostatin partially inhibited cell death induced by JL5. All experiments were performed at least 3 times with similar results except (c), which was performed twice with similar results