P lkb1

Rabbit polyclonal antibodies specific for ICAM-1, p-AMPK, AMPK, p-LKB1, LKB1, p-CaMKII, CaMKII, p-c-Jun, c-Jun, and β-actin, anti-mouse and anti-rabbit IgG-conjugated horseradish peroxidase, and Protein A/G beads were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Compound C and adenosine-9-β-d-arabino-furanoside (AraA) were purchased from Calbiochem (San Diego, CA). Human full-length adiponectin was purchased from R&D Systems (Minneapolis, MN). The AP-1 luciferase plasmid was purchased from Stratagene (La Jolla, CA). The pSV-β-galactosidase vector and luciferase assay kit were purchased from Promega (Madison, MA). All other chemicals were purchased from Sigma-Aldrich (St. Louis, MO).

Western blotting was performed as previously described [3 (link)]. Briefly, the cells were lysed with RIPA buffer (Sigma Aldrich) containing 150 mM NaCl, 1.0% Nonidet-P 40, 0.5% sodium deoxycholate, 0.1% sodium dodecyl sulfate, 50 mM Tris (pH 8.0), a protease inhibitor cocktail, and PhoSTOP (Roche Molecular Biochemicals, Basel, Switzerland). All antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA), and antibodies against VE-cadherin, p-LKB1, and LKB1 were purchased from Santa Cruz Biotechnology (Santa Cruz, California, USA). Secondary antibodies (anti-rabbit IgG or anti-mouse IgG, 1:2000) were purchased from Cell Signaling Technology. Image densities were quantified using the National Institutes of Health ImageJ 1.41q software.