LPS Regulates SOCS2 Transcription in a Type I Interferon Dependent Autocrine-Paracrine Loop
Figure 2
Type I IFN regulated IRFs, but not NF-κB were required for SOCS2 mRNA induction by TLR4 signaling in human moDCs.
(A) NF-κB, IRF1 and IRF3 nuclear translocation in TLR4 signaling in human moDCs. moDCs were stimulated with LPS at the indicated time points. Nuclear protein was extracted for Western Blotting measurement. The levels of p65NF-κB, IRF1, IRF3 and Lamin A (as loading control) were detected. (B–D) The effect of silencing NF-κB, IRF1 and IRF3 on the induction of SOCS2 mRNA in LPS stimulated human moDCs. moDCs were transfected with (B) NF-κB, (C) IRF1, (D) IRF3 siRNAs or control siRNA and incubated for 24 hours. The transfected cells were exposed to LPS for 0 h, 8 h or 24 h, the cells were then harvested for the mRNA measurement by qRT-PCR to demonstrate the effect of SOCS2 mRNA induction (left part) and efficiency of target gene knockdown (right part). Data shown are representative of three independent experiments and are expressed as the –fold induction of the gene of interest at different time points compared to time point 0 for control siRNA where the values were set to 1. Statstical significance comparing results from the target siRNA to control siRNA group is indicated (* p<0.05; ** p<0.01).
