Phospho p44 p42 erk thr202 tyr204

RAW 264.7 cells were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA, TIB-71). Auraptene (AU, >98%, Figure 1(a)) was purchased from ChemFaces Biochem, Wuhan, Hubei, China. Sigma (St Louis, MO, USA) supplied potassium ferricyanide, ferric chloride, and dimethyl sulfoxide (DMSO). Santa Cruz Biotechnology (Dallas, TX, USA) supplied anti-iNOS and COX-2 polyclonal antibodies (pAb). We purchased antibodies against TNF-α, phospho-p38 MAPK Thr180/Tyr182, phospho-c-JNK (Thr183/Tyr185), phospho-p44/p42 ERK (Thr202/Tyr204), phospho-IκBα Ser32/36, and phospho-NF-κB p65 (Ser536) pAbs from Cell Signaling (Beverly, MA, USA). Anti-IL-1β and anti-HO-1 pAbs were purchased from BioVision (Milpitas, CA, USA) and Enzo (Farmingdale, New York, USA), respectively. The antibody against α-tubulin was purchased from NeoMarkers (Fremont, CA, USA). AU was dissolved in 0.1% DMSO.

Columbianadin (CBN, >98%) was purchased from ChemFaces Biochem, Wuhan, Hubei, China. Dimethyl sulfoxide (DMSO), PD98059, SP600125, BAY11-7082, and DMPO were purchased from Sigma (St Louis, MO, USA). The antibodies against phospho-p38 MAPK Ser¹⁸² (pAb), IκBα (44D4), phospho-c-JNK (Thr¹⁸³/Tyr¹⁸⁵), p38 MAPK, NF-κB p65 (mAb), phospho-p44/p42 ERK (Thr²⁰²/Tyr²⁰⁴), and phospho-NF-κB p65 (Ser⁵³⁶) pAb were all purchased from Cell Signaling (Beverly, MA, UAS). Anti-HO-1 pAb was purchased from Enzo (Farmingdale, New York, USA). The monoclonal antibody against α-tubulin was derived from NeoMarkers (Fremont, CA, USA). CBN was dissolved in 0.1% DMSO. All other chemicals and reagents used in this study were commercially purchased from Sigma until specified otherwise.