Fig. 1. 10d and 10e induce apoptosis in HL-60 cells associated with upregulation of Noxa and downregulation of Mcl-1 and c-Flip.
A The chemical structures of 10d and 10e. B HL-60 cells were treated with 10d and 10e at the indicated concentrations for 6 h and the cells in subG1 phase were determined using FACS analysis after staining with PI. con, untreated cells. C HL-60 cells treated with 10d or 10e at different concentrations for 6 h, the relative levels of each protein were analyzed with western blotting using specific antibodies. D The HL-60 cells were treated with 1 μM 10d or 2 μM 10e for the indicated time, the levels of PARP, c-FlipL, Noxa proteins were examined. E HL-60 cells were pretreated with 100 μM Z-VAD-FMK or 25 μM q-VD-OPH for 1 h, following by treatment with 2 μM 10e for 6 h, the relative levels of each protein were analyzed with western blotting using specific antibodies. F HL-60 cells were pretreated with 10 μM MG132 for 1 h and then treated with or without 2 μM 10e for 4 h. The relative levels of PARP, c-Flip, Mcl-1, and Noxa were determined using Western blot analysis. The cell lysates was also immunoprecipitated with an anti-Mcl-1 antibody, and then immunoblotted with an anti-Mcl-1 antibody and an anti-ubiquitin antibody