Figure 1. A) Representative Western blot image showing the presence of p50 subunit of NF-B in the nuclear extracts prepared from HASM cells. C: control cells; TNF-: cells treated with 50 ng/ml TNF- for 22 h; SN50: cells pretreated with 18 µM SN50 followed by 50 ng/ml TNF- for 22 h; and SN50M: cells pretreated with 18 µM SN50M followed by 50 ng/ml TNF- for 22 h. Note decreased amount of p50 in nuclear extracts prepared from SN50-treated cells indicating decreased NF-B activation. Nuclear actin is shown as an internal control; n = 3. B) Results of quantitative real-time PCR shown as fold change in CD38 mRNA expression in HASM cells. Note significant (P<0.05) attenuation of CD38 expression in cells pretreated with SN50; n = 3. C) Results of transient transfection using the various IB mutants on TNF--induced CD38 expression in HASM cells. Lanes 1 and 2: untreated and TNF--treated cells after electroporation; lanes 3 and 4: cells transfected with the vector alone; lanes 5 and 6: cells transfected with the N-terminal deletion mutant; lanes 7 and 8: cells transfected with the C-terminal deletion mutant; and lanes 9 and 10: cells transfected with the IKK dominant negative mutant. Note attenuation of TNF--induced CD38 expression in the presence of N-terminal deletion and IKK dominant negative mutants and increased expression in the presence of the C-terminal deletion mutant. Representative of 3 experiments.