Phospho jnk t183 y185

Immunoblotting was performed as previously described^{4 (link)}. Signal was visualized by chemiluminescence on X-ray films or by digital acquisition on a ChemiDoc MP (Biorad). Densitometric quantification of Western blot results was performed using ImageJ 2.0.0-rc-49/1.51d (NIH). Antibodies used were phospho-T202/Y204 ERK (D13.14.4E; Cell Signaling Technologies), ERK (Cell Signaling Technologies), phospho-T183/Y185 JNK (81E11; Cell Signaling Technologies), phospho-T183/Y185 JNK (98F2; Cell Signaling Technologies), JNK1/2/3 (Cell Signaling Technologies), JNK (Sigma), phospho-T180/Y182 p38 (D3F9; Cell Signaling Technologies), p38α (Cell Signaling Technologies), GAPDH (Millipore), NeuN (Abcam), GFAP (Abcam).

Defatted soy protein and fat-free casein were procured from Sakthi Sugars Limited, Coimbatore, India, and Sisco Research Laboratory, Mumbai, India, respectively. Insulin and glucose assay kits were obtained from Monobind Microwells Inc, CA, USA, and Agappe Diagnostics Pvt. Ltd, Kerala, India, respectively. Antibodies such as phospho (T183/Y185) JNK and phospho-IKKα/β (Ser176/180) and anti-IKKβ were purchased from Cell Signaling Technology, MA, USA. Anti-JNK was obtained from Santacruz Biotechnology, CA, USA. Enhanced chemiluminescence (ECL, Immobilon HRP Western Substrate) kit was purchased from Thermo Scientific, MA, USA. Nuclear extraction kit and NF-κB p65 transcription factor assay kit were bought from Cayman Chemical Company, MI, USA. Supersensitive polymer-horseradish peroxidase immunohistochemistry detection kit was purchased from Biogenex laboratories, San Ramon, CA, USA. Antibodies, namely, anti-3-NT and anti-4-HNE, were purchased from Invitrogen, USA, and Merck (Calbiochem), Darmstadt, Germany, respectively. Primers were purchased from Sigma-Aldrich, MO, USA, and the SYBR Green-qPCR master mix was purchased from Thermo Scientific, MA, USA.