Figure 4

Effects of antisense oligonucleotide against NF-κB, curcumin and anti-TNF-β on IL-1β-induced phosphorylation and translocation of NF-κΒ-p65 in nuclear extracts in human chondrocytes. (1) Serum-starved primary human chondrocytes were either left untreated or were treated with 10 ng/ml interleukin 1β (IL-1β), 5 μM curcumin, 0.5 μM sense oligonucleotide (SO) control or antisense oligonucleotide (ASO) against nuclear factor κB (NF-κB) in the presence of 10 μl/ml Lipofectin transfection reagent or 10 ng/ml anti-tumor necrosis factor β (anti-TNF-β) alone for 24 hours; or (2) cells were pretreated with 10 ng/ml IL-1β for 1 hour, followed by cotreatment with 5 μM curcumin, 0.5 μM SO control or ASO against NF-κB in the presence of 10 μl/ml Lipofectin transfection reagent or 10 ng/ml anti-TNF-β for 24 hours. Nuclear cell fractions were prepared and examined by Western blot analysis using antibodies against phospho-specific p65 and nuclear protein poly(ADP-ribose) polymerase as a loading control. Western blots shown are representative of three independent experiments.