Figure 8

Activation of NF-κB by BA is important for increasing apoptosis/cell death in combination with 2ME2 or Doc. A. DAPI analysis showing that LN-AI/dnI clones 7 and 20 undergo less apoptosis (% condensed fragmented nuclei) when treated with MB or DB for 24 h compared to LN-AI/neo negative control (N). n = 6, three independent experiments; *, P < 0.04. Western blot analysis showing less cleaved PARP in LN-AI/dnI clones 7 and 20 compared to LN-AI/neo (N) cells treated with MB or DB for 24 h. B. Trypan blue exclusion and Western blot analysis showing that knockdown of p65 in LNCaP/shp65-2 and DU145/shp65-1 lowers cell death (n = 6, three independent experiments; *, P < 0.003) and cleaved PARP when treated with MB or DB and compared to control (C) treated LNCaP/shGFP and DU145/shGFP cells. C. Trypan blue exclusion assay showing that DU145 cells pretreated (PT) with NF-κB inhibitor parthenolide (Pa; 10 μM) for 24 h then treated (T) with MB or DB + Pa for 48 h underwent less cell death (% dead cells) compared to cells not pretreated with Pa and treated with MB or DB. Also shown are cells pretreated and treated with Pa alone. n = 6-7, four independent experiments. *, P < 0.002. D. Trypan blue exclusion assay showing increase in cell death in DU145 and PC3 cells treated for 72 h with 2ME2 + BA (MB) or Doc + BA (DB) compared to BA (B; 10 μM), 2ME2 (M; 5 μM), Doc (D; 1 nM) alone and control cells. n = 5-6, three independent experiments; *, P < 0.004.