Figure 3

IKK activation is required for the induction of miR-425 in response to IL-1β induction. (A) AGS cells were treated with IL-1β in the presence of the IKK inhibitor TPCA-1, the p38 MAPK inhibitor BIX02188 and the JNK inhibitor SP600125. Mature miR-425 expression at 12 h after treatment was quantitated by real-time PCR. The fold change of relative miR-425 expression (miR-425/U6) was normalized to that observed in PBS-treated cells. (B) The expression of primary miR-425 (pri-miR-425) was analyzed by real-time PCR as in A. (C) The expression levels of NF-kappaB protein and pri-miR-425 were analyzed by real-time PCR in AGS cells treated with siRNAs for NF-kappaB. (D) AGS cells were treated with chemistry inhibitors or siRNAs for NF-kappaB. Cell lysates were obtained 24 h after IL-1β treatment and immunoblotted with PTEN antibodies. (E) Western blot analysis of phosphorylated NF-kappaB p65 (serine 536). (F) The expression levels of NF-kappaB protein and pri-miR-425 were analyzed by real-time PCR in NCI-N87 cells treated with siRNAs for NF-kappaB. (*p < 0.05; **p < 0.01; ***p < 0.001).