Anti phospho p38 9211

Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS) were obtained from Life Technologies (Tokyo, Japan). EPA, DHA, and 4-HHE were purchased from Cayman (Ann Arbor, MI, USA). Anti-β-actin antibody (A5316) and N-acetyl-l-cysteine (NAC) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Fatty acid-free bovine serum albumin (BSA) was purchased from Nacalai Tesque (Kyoto, Japan). Anti-epoxide hydrolase 2 (sEH) antibody was purchased from Gene Tex Inc. (Irvine, CA, USA). Anti-p38 kinase (p38) (#9212) and anti-phospho-p38 (#9211) antibodies were purchased from Cell Signaling (Danvers, MA, USA). Anti-PECAM-1 (M-20) antibody and anti-heme oxygenase 1 (HO-1) antibody were purchased from Santa Cruz (Dallas, TX, USA). Anti-cytochrome P450 2J2 (CYP2J2) antibody was purchased from Origene (Rockville, MD, USA). Horseradish peroxidase (HRP)-conjugated anti-mouse and anti-rabbit antibodies were purchased from Amersham Biosciences Corp. (Piscataway, NJ, USA). Small interfering RNA (siRNA) was purchased from Life Technologies (Tokyo, Japan). SB203580 was purchased from Calbiochem (Cambridge, UK). All other reagents and chemicals were from standard suppliers.

Total protein was extracted from the cells with protein lysis buffer, and the protein concentration was subsequently measured by Lowry protein assay. Twenty micrograms of total protein from each sample was resolved by 10% or 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), transferred to a polyvinylidene difluoride membrane (Millipore), blocked in 5% nonfat milk and immunoblotted with various primary antibodies and horseradish peroxidase-conjugated secondary antibodies. Anti-phospho ERK1/2 (9101, 1:1000), anti-ERK1/2 (9102, 1:500), anti-phospho RelA (3033, 1:1000), anti-phospho p38 (9211, 1:500), and anti-p38 antibodies (9212, 1:500) were purchased from Cell Signaling Technology. An anti-cyclin E1 (11554-1-AP, 1:500) antibody was purchased from Proteintech. An anti-GAPDH (KC-5G5, 1:5000) antibody was purchased from Kangchen. An anti-actin antibody (A5441, 1:2000) was purchased from Sigma-Aldrich. Antibody binding was detected using SuperSignal West Pico Chemiluminescent Substrate (Pierce).

Untreated Bioflex® plates (BF-3001U) and laminin Bioflex® plates (BF-3001L) were purchased from Dunn Labortechnik GmbH (Asbach, Allemagne). Poly-L-lysine (P9404), propidium iodide (P4170), Cell Lysis Reagent (C2978), TRITC-phalloidin (P1951), Hoechst (H2261), and anti α-tubulin antibody (T6074) were purchased from Sigma-Aldrich (Lyon, France). Laminin (23017-015), BME (41010-026), DMEM (11960), HBSS (14060-040), Pierce®ECL 2 Western Blotting substrate kit (PI80196), Alexa 488 goat anti-mouse IgG (A21121), anti-APP antibody (51-2700), and CM-H2DCFDA probe (C6827) were purchased from Thermo Fisher Scientific (Courtaboeuf, France). Cy3 goat anti-rabbit (111-165-003) was purchased from Jackson ImmunoResearch (Cambridge, UK). Anti-MAG (MAB1567), anti-MBP (MAB381), anti-phospho-ERK1/2 (05-797R), and anti ERK1/2 (06-182) antibodies were purchased from Millipore (Molsheim, France). Anti-SMI312 (837904) antibody was purchased from Biolegend (San Diego, USA). Anti-Caspr (ab34151) and rabbit polyclonal β-actin (ab8227) antibodies were purchased from Abcam (Cambridge, UK). Anti-PDGFR-α antibody (sc-338) was purchased from Santa Cruz (Dallas, USA). Anti-PLP antibody (NB10074503) was purchased from Novus Biologicals. Anti-JNK (9252), anti-phospho-JNK (9251S), anti-p38 (212), and anti-phospho-p38 (9211) antibodies were purchased from Cell Signaling (Danvers, USA).