Figure 5.

NLRP11 silencing enhances pro-inflammatory responses. siRNA-mediated knockdown of NLRP11 in PMA-differentiated (PMA diff.) THP1 cells. Mean + S.D. of two independent experiments, each conducted in biological triplicates (n = 6), is shown. A, IL-8 release from THP1 cells transfected with an NLRP11-specific siRNA or a non-targeting control siRNA (siRNA ctrl) treated for 16 h with LPS or TNF, respectively. B, IFNβ release from THP1 cells transfected with a NLRP11-specific siRNA or a non-targeting control siRNA treated for 16 h with SeV. Ctrl, non-treated cells; n.d., not detectable C, end point PCR with NLRP11 and GAPDH-specific primer sets of a representative experiment from A. D, NF-κB activation in THP1blue reporter cells measured by SEAP secretion upon the indicated siRNA treatment. Cells were stimulated for 16 h with 100 ng/ml LPS or 50 ng/ml TNF, respectively. E, immunoblotting of THP1blue cells treated for the indicated time with 100 ng/ml LPS. Phosphorylation of p38 and total p38 as a loading control are probed. One representative blot of three experiments is shown. ***, p < 0.0005 (two way ANOVA with Sidak's multiple comparisons test).