Rpmi complete rpmi 1640 medium

Monocyte isolation kits were from Miltenyi Biotec (Auburn, CA, USA). RPMI Complete (RPMI 1640 medium supplemented with FBS, 2-mercaptoethanol, sodium pyruvate, L-glutamine and HEPES) was purchased from Invitrogen Life Technologies (Carlsbad, CA, USA). Ultrapure LPS from Escherichia coli 0111:B4 was purchased from Sigma-Aldrich (St. Louis, MO, USA). Anti-phospho-NF-κB p65 (Ser536), pGSK3β (Ser9), pCREB (Ser133), pERK1/2 (Thr202/Tyr204), pSAPK/JNK (Thr183/Tyr185), p-p38 MAPK (Thr180/Tyr182), GAPDH, β-actin and goat anti-rabbit IgG-HRP Abs, as well as non-targeted Signal Silence Control siRNA were from Cell Signaling Technology (Danvers, MA, USA). Anti-pSGK (Ser422) goat Abs came from Santa Cruz Biotechnology (Santa Cruz, CA, USA), while mouse anti-goat IgG-HRP Abs were from Rockland Immunochemicals (Limerick, PA, USA). MaR1, RVD1 and RVD2 were purchased from Cayman Chemical (Ann Arbor, MI, USA). Enhanced chemiluminescence kits were purchased from Thermo Scientific (Rockford, IL, USA). Cytokine ELISA kits (TNF, IL-10, IL-8 and IL-1β) were from eBioscience (San Diego, CA, USA). IL-12/23 p40 ELISA kits came from Boster Immunoleader (Pleasanton, CA, USA). The plasmids HA pcDNA3 and HA pcDNA3-GSK3β(S9A), were gifts from Jim Woodgett (Addgene; plasmids 10792, 14754). CREB siRNA was purchased from Dharmacon (Lafayette, CO, USA).

Monocyte isolation kits came from Miltenyi Biotec (Auburn, CA). RPMI Complete (RPMI 1640 medium supplemented with 10 % FBS, 50 μM 2-ME, 1 mM sodium pyruvate, 2 mM l-glutamine, 20 mM HEPES, 50 U/ml penicillin, and 50 μg/ml streptomycin) was purchased from Invitrogen Life Technologies (Carlsbad, CA). Ultrapure LPS from E. coli 0111:B4 and other human TLR agonists were purchased from InvivoGen (San Diego, CA). 2-Amino-4-(3,4-(methylenedioxy)benzylamino)-6-(3-methoxyphenyl)pyrimidine was bought from Calbiochem (San Diego, CA). All antibodies came from Cell Signaling Technology (Danvers, MA). Enhanced chemiluminescence kits were purchased from Thermo Scientific (Rockford, IL). Cytokine ELISA kits (TNF, IL-6 and IL-12/23 p40) were purchased from eBioscience (San Diego, CA). The Wnt pathway inhibitors, N-(6-methyl-2-benzothiazolyl)-2-[(3,4,6,7-tetrahydro-4-oxo-3-phenylthieno[3,2-d]pyrimidin-2-yl)thio]-acetamide (IWP-2) and benzoic acid, 2-phenoxy-,2-[(5-methyl-2-furanyl)methylene] hydrazide (PNU-74654), came from Tocris Biosciences (Minneapolis, MN). The GSK3 inhibitor, SB216763, was bought from Sigma-Aldrich (St. Louis, MO). Human Monocytes Nucleofector kit, including pCMV-GFP, came from Amaxa AG (Koln, Germany), while SMARTpool siRNA came from Dharmacon (Lafayette, CO).