Fig. 6.
DIM induces apoptosis and enhances HDAC inhibitor-induced apoptosis. (A) HT-29 cells were treated with various doses of DIM for 24h. Apoptosis was analyzed using the Cell Death Detection ElisaPLUS kit as described in Materials and Methods. (B) HT-29 cells were treated with or without 20 μM of DIM for 24 hours, followed by various doses of SAHA or Trichostatin A (TSA) for additional 24h. Apoptosis was analyzed as described in (A). The average results from three independent experiments were shown. (C) DIM/SAHA treatment induced Bak activation. HT-29 cells were treated with 20 μM DIM for 24h, 2 μM SAHA for 24h, or 20 μM DIM for 24h followed by 2 μM SAHA for additional 24h. Bak dimerization was detected by cross-linking experiments as described in Materials and Methods. Protein lysates were analyzed by western blotting. (D) HT-29 cells were treated as described in (C) and western blot was performed using the indicated antibodies.