Irak1 d51g7

Prostate cancer cell lines PC3, DU145, and LNCaP were obtained from the American Type Culture Collection (Manassas, VA). Cell lines were authenticated by examination of morphology and growth characteristics and confirmed to be mycoplasma-free. Cells were maintained in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% FBS (Life Technologies, Grand Island, NY), and cultured for less than 6 months. For western blotting, the specific primary antibodies indicated were used to detect the following proteins: FOXP3 (ab450, Abcam, Cambridge, MA), Foxp3 (hFOXY, eBioscience, San Diego, CA), NF-κB p65 (D14E12, Cell Signaling, Danvers, MA), IRAK1 (D51G7, Cell Signaling), and TRAF6 (D21G3, Cell Signaling), Irak1 (H-273, Santa Cruz Biotechnology), and Traf6 (H-274, Santa Cruz Biotechnology). For immunohistochemistry (IHC), the indicated specific antibodies were used to detect the following mouse proteins: Foxp3 (Poly2638b, BioLegend, San Diego, CA), NF-κB p65 (D14E12), Irak1 (H-273), and Traf6 (H-274). Scramble control or miR-146a/binhibitors (Life Technologies) were transfected into PC3 and DU145 cells. Lipopolysaccharide (LPS) (O111B4, Sigma, St. Louis, MO) and bortezomib (Selleck Chemicals, Houston, TX) were used for intraperitoneal injection in mice.