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. Author manuscript; available in PMC: 2012 Dec 10.
Published in final edited form as: Biochim Biophys Acta. 2007 Jul 4;1772(9):1103–1111. doi: 10.1016/j.bbadis.2007.06.004

Fig. 6.

Fig. 6

Chemotherapy-HDI combinations increase pro-caspase 8 activation. A, MDA-MB-231 cells were untreated (lane 1), or treated with 1 μM doxorubicin (lane 2), 5 mM HDI (sodium butyrate, lane 3), or doxorubicin plus HDI (lane 4) for 48 hours. Western blots were analyzed for (A) clusterin, (B) pro-caspase 8, (C) IκBα, (D) FasL, (E) FADD, (F) RIP and tubulin as indicated. Fold changes in protein expression/tubulin ratios, relative to untreated cells, are indicated below each panel. G, Cells were treated as for part A, and caspase 8 activity was measured in the lysates using the chromogenic substrate Ac-IETD-pNA (N-acetyl-Ile-Glu-Thr-Asp p-nitroanilide). The analysis was repeated in triplicate, and fold change in activity relative to untreated cells is shown. H, MDA-MB-231 cells were transfected with oligonucleotide RNAi duplexes targeting a control sequence (lanes 1–4) or clusterin (lanes 5–8). Cells were left untreated (lanes 1 and 5) or were treated with 1 μM doxorubicin (lanes 2 and 6), HDI (5 mM sodium butyrate, lanes 3 and 7), doxorubicin-HDI combined (lanes 4 and 8). Western blots were analyzed for clusterin, pro-caspase 8 and tubulin as indicated. The results show that caspase 8 is activated following doxorubicin-HDI treatment, and that clusterin does not affect caspase 8 cleavage.