Figure 6.

The effect of RNAi-mediated inhibition of transcription factor genes in J77A4.1 macrophages on IL-6 (A) and TNF-α (B) production in response to LPS stimulation. Pools of four siRNA duplexes per gene (2 μM concentration) were transfected into J77A4.1 cells, LPS was added, and cytokine production was monitored as described in Materials and Methods. Shown are the results for four negative controls (nontargeting siRNAs from Dharmacon; all data are normalized to the first negative control), two positive controls (TLR4 and IL-6), and 15 transcription factors identified by our analysis of gene expression data. To verify that the phenotypes observed were caused by knockdown of that gene, the four siRNA duplexes were transfected individually for the seven genes that had the strongest effect, and IL-6 production was monitored (C). At least two independent siRNAs induced a phenotype for each gene. In all three panels, plotted are means of three independent measurements with error bars representing SD.