Figure 5.

Loss of RNF40 and RNF20 reduces induction of NF-κB target gene expression upon TNFα treatment. HCT116 siControl, siRNF40, siRNF20, and siWAC cells were stimulated with 10ng/mL of TNFα for 0, 0.5, and 6 h in three independent experiments, 48 h after siRNA transfection. [A] The siRNA-mediated knockdowns were verified, and relative mRNA expression levels of RNF40, RNF20, and WAC were not affected by TNFα treatment [n = 3]. Mean ± SEM. [B] Relative gene expression of several NF-κB target genes was assessed using qRT-PCR [n = 3]. Generally, the knockdown of RNF40 or RNF20 resulted in a reduced induction of these genes upon TNFα treatment, especially after 30 min, whereas WAC depletion only showed mild effects compared with the control siRNA. Mean ± SEM. SEM, standard error of the mean; qRT-PCR, quantitative real-time polymerase chain reaction.