n-3 DPA (purity ≧ 98%) solution in 100% ethanol was obtained from Cayman Chemical (Ann Arbor, MI, USA). n-6 DPA (purity > 99%) was purchased from Nu-Chek Prep, Inc. (Elysian, MN, USA). DPA was dissolved in 100% ethanol to make a 30 mM stock solution. The stock solutions were stored at −20 °C, then diluted to final concentrations before use. Sphingosylphosphorylcholine (SPC) was purchased from Biomol (Plymouth Meeting, PA, USA). Fura-2/AM was purchased from Dojindo (Kumamoto, Japan). Bradykinin (BK) was purchased from Peptide institute, Inc. (Osaka, Japan). All other chemicals were purchased from Katayama Chemical (Osaka, Japan).
The following antibodies were used: anti-phospho-MYPT1 (Thr850) with phosphorylation site corresponding to Thr853 in human MYPT1 (Upstate), anti-GAPDH monoclonal antibody (Wako), anti-phospho-myosin light chain 2 (Ser19) monoclonal antibody (Cell Signaling), anti-myosin light chain (20 kDa) monoclonal antibody (Sigma Aldrich), anti-Rho-kinase-2 polyclonal antibody (H85, Santa Cruz), and anti-rabbit Alexa Fluor 488 (Thermo Fisher). Secondary HRP-labeled antibodies (anti-mouse and anti-rabbit) were purchased from Promega.
The following antibodies were used: anti-phospho-MYPT1 (Thr850) with phosphorylation site corresponding to Thr853 in human MYPT1 (Upstate), anti-GAPDH monoclonal antibody (Wako), anti-phospho-myosin light chain 2 (Ser19) monoclonal antibody (Cell Signaling), anti-myosin light chain (20 kDa) monoclonal antibody (Sigma Aldrich), anti-Rho-kinase-2 polyclonal antibody (H85, Santa Cruz), and anti-rabbit Alexa Fluor 488 (Thermo Fisher). Secondary HRP-labeled antibodies (anti-mouse and anti-rabbit) were purchased from Promega.