Figure 3

TNFα induced midkine expression through NF-κB pathway. A. LNCaP cells were not treated or treated for 48 h with increasing dosages of TNFα, without (top blot) or with 18 μM NF-κB inhibitor (bottom blot); 20 μl of each medium supernatant was subjected to Western blot analysis of midkine expression. B. Densitometry of A; the untreated control group was arbitrarily assigned a value of 1; the integrated density values (IDVs) of the protein bands from other groups were divided by that of the control group (i.e., ratio of IDVs) to represent the relative MDK levels; solid line, TNFα only; dotted line, TNFα with 18 μM NF-κB inhibitor. Data presented were average ± standard deviations (error bars) of three independent experiments. C. LNCaP cells were not treated or treated for 48 h with 20 ng/ml TNFα, and without or with increasing dosages of NF-κB inhibitor; 20 μl of each medium supernatant was subjected to Western blot analysis of midkine expression. D. Densitometry of C as described in B. Data presented were average ± standard deviations (error bars) of three independent experiments.