Fig. 3.

Nur77-active C-DIMs induce apoptosis in UC-5 and KU7 cells and also act through nuclear Nur77. A, effects of leptomycin B on PARP cleavage. UC-5 cells were treated for 24 h with DIM-C-pPhOCH₃ alone or in the presence of leptomycin B (0.05 ng/ml), and Western blot analysis of nuclear and cytosolic cell lysates was used to detect Nur77 and PARP cleavage. Sp1, loading control for nuclear fraction; N.S., nonspecific band for loading control. B, immunostaining for Nur77. UC-5 cells were treated with DMSO or 15 μM DIM-C-pPhOCH₃ for 24 h and immunostained with IgG or Nur77 antibodies. C, dose-dependent induction of apoptosis by C-DIMs. UC-5 and KU7 cells were treated with DMSO or different concentrations of DIM-C-pPhOCH₃ or DIM-C-Ph for 48 h. D, inhibition of Nur77-active C-DIMs-induced apoptosis by Z-VAD-fmk. UC-5 cells were treated with Nur77-active C-DIMs alone or in combination with 10 μM Z-VAD-fmk, and whole-cell lysates were analyzed by Western blot analysis.