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. Author manuscript; available in PMC: 2010 Jul 9.
Published in final edited form as: Cancer Prev Res (Phila). 2009 May 26;2(6):581–589. doi: 10.1158/1940-6207.CAPR-08-0142

Figure 4. Survivin plays a key role in DIM/butyrate-induced apoptosis.

Figure 4

(A) Overexpression of survivin blocked DIM/butyrate-induced apoptosis. HT-29 cells were transfected with 0.5 μg of phrGFP or phrGFP-survivin vectors in 6-well plates. 24h after transfection, cells were treated with 40 μM DIM for 24h followed by various concentrations of butyrate for 24h. Expression of hrGFP-survivin was confirmed by western blotting with anti-hrGFP antibody. Apoptosis was analyzed by DAPI staining as described in Materials and Methods. The average result from three independent experiments was shown. (B) Knocking-down of survivin by siRNA sensitized HT-29 cells to butyrate-induced apoptosis. Survivin targeting siRNA and negative control siRNA were transfected into HT-29 cells as described in Materials and Methods. Down-regulation of survivin was confirmed by western blotting of protein samples collected at 48h after transfection. At 24h post siRNA transfection, various concentrations of butyrate were added to cells and apoptosis was analyzed 24h after butyrate exposure by TUNEL assay. The average result from three independent experiments was shown.