Figure 4
From: NF-κB activation enhances cell death by antimitotic drugs in human prostate cancer cells
Inhibition of NF-κB with dominant negative IκBα (dnI) or p65 shRNA decreases apoptosis induced by 2ME2 and Doc. A. DAPI analysis showing that LN-AI/dnI clones 7 and 20 undergo less apoptosis (% condensed fragmented nuclei) when treated with 2ME2 (M; 5 μM) or Doc (D; 1 nM) for 48 h compared to LN-AI/neo negative control (N). n = 8, four independent experiments; *, P < 5 × 10-5. B. Western blot analysis showing less cleaved PARP in LN-AI/dnI clones 7 and 20 compared to LN-AI/neo (N) cells treated with M or D for 48 h. No cleaved PARP was detected in control (Cont) treated cells. Endogenous phospho (P)-IκBα increases in LN-AI/dnI-7 and neo (N) cells but not in LN-AI/dnI-20 cells when treated with M or D for 24 h. Total levels of endogenous IκBα decrease in all cells treated with M or D compared to Cont treated cells. Transfected (tx) dnI protein (migrates slightly faster than endogenous IκBα) remains in LN-AI/dnI clones 7 and 20 but is not detected in LN-AI/neo cells. ns, non-specific band. Coomassie blue stain of total protein is loading control. C. DAPI and Western blot analysis showing less apoptosis and cleaved PARP in p65 knockdown LNCaP/shp65-2 (p65) compared to control LNCaP/GFP (C) cells treated with 2ME2 or Doc for 72 h. n = 6, three independent experiments; *, P < 5 × 10-3.