FIGURE 1.

SET7/9 knockdown inhibits expression of key NF-κB-dependent genes. A, HEK293 cells were transiently transfected with plasmid vectors expressing SET7/9 shRNA (SI) or scrambled sequence (CON). Inflammatory gene expression was determined by RT-PCR using 18S primers as internal control after cells were stimulated with or without TNF-α (10 ng/ml) for various time periods. B, cell lysates were immunoblotted (IB) with SET7/9 and β-actin antibodies to confirm the knockdown of SET7/9 protein. C, THP-1 cells were stably transfected with lentiviral vectors expressing SET7/9 shRNA or scrambled sequence, and RT-PCRs were performed to detect MCP-1, TNF-α, and IL-8 expression after TNF-α treatment (10 ng/ml) for various time periods. D, Western blot to demonstrate SET7/9 knockdown efficiency by the lentiviral vector in THP-1 cells. E, quantification of TNF-α-induced gene expression at the 1 h time point in THP-1 cells. ^*, p < 0.001 SI + TNF-α versus CON + TNF-α, n = 3.