Anti rsk

All of the drugs, including urethane, fructose, resveratrol, compound C, and dimethyl sulfoxide (DMSO), and the mouse anti-actin, goat anti-rabbit, and goat anti-mouse IgG secondary antibodies were obtained from Sigma-Aldrich (Sigma Chemical Co., St. Louis, MO, USA). Anti-p-AMPK^T172, anti-AMPK, anti-ACC^S79, anti-p-ERK^T202/Y204, anti-ERK, anti-nNOS^S1416, anti-nNOS, anti-eNOS^S177, anti-eNOS, anti-iNOS, anti-p-RSK^T359/S363, and anti-RSK antibodies were purchased from Cell Signaling Technology (Beverly, MA, USA). Anti-p22-phox and anti-nitrotyrosine were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-p47-phox and anti-p67-phox were purchased from Millipore (Bedford, MA, USA). Anti-Cu/Zn-SOD and anti-Mn-SOD were obtained from StressGen Biotechnologies (La Jolla, CA, USA) and Abcam (Cambridge, UK), respectively.

Protein was detected using primary antibodies detecting YBX1, CDK6, TCTP, GAPDH, H3, and pYBX1 (ab138654 and ab138654 detect endogenous levels of YB1 only when phosphorylated at S102), pAKT, pERK, pRSK, AKT, ERK, RSK, PDGFRB, PCK2, and BAD. Anti-pAKT, anti-pERK, anti-pRSK, anti-AKT, anti-ERK, and anti-RSK were purchased from Cell Signaling Technology (CST) and all the other antibodies were purchased from Abcam. Western blotting was performed as in previous studies (Li et al, 2016 (link)). Briefly, protein samples collected above were separated on SDS–PAGE gels via electrophoresis, transferred to PVDF membranes (Millipore), blocked, and probed using appropriate primary antibodies. Samples were then probed with appropriate secondary antibodies and protein bands were detected using the ECL reagent (GE Healthcare). GAPDH was used as a loading control for total and cytoplasmic protein, whereas H3 was used for nuclear protein. ImageJ was used to quantify protein band densitometry.

This assay was performed as described previously [10 (link)]. The following antibodies were used in this study: Anti-Irf6, anti-Blnk, anti-ErbB2, anti-RSK, and anti-phospho-RSK (all from Cell Signaling Technology, Danvers, MA, USA), anti-ΔNp63 and anti-TAp63 (BioLegend, San Diego, CA, USA), anti-CDK4 (Santa Cruz Biotechnology, Dallas, TX, USA), and anti-β-actin (Santa Cruz Biotechnology and Sigma-Aldrich).