Western blot was performed as previously described20. Whole cell lysates from cultured cells are were harvested in cell lysis buffer containing 20 mM Tris-HCl (pH 7.5), 150 mM NaCl, 1% Triton X-100, 1 mM EDTA, 1 mM EGTA, 2.5 mM sodium pyrophosphate, 1 Mm β-Glycerolphosphate, 50 mM NaF, 1 mM Na3VO4 and supplemented with protease inhibitor cocktail (#P8340, Sigma-Aldrich) for 30 min at 4 °C. Then the cell lysates were centrifuged at 4 °C (12, 000 rpm) for 15 min and protein extract supernatant was collected. Protein concentrations were determined with the Bradford protein assay kit (#500-0006, Bio-rad) using a Beckman DU-800 spectrophotometer (Fullerton, CA). Total cell lysates (20–30 μg) were separated by SDS-PAGE and transferred to nitrocellulose membrane (Bio-rad).The membranes were then blocked with diluted Odyssey® blocking buffer (#927-40000, LI-COR Biosciences, Lincoln, NE) for 1 h. The membranes were incubated with appropriate primary antibodies overnight at 4 °C and then washed with 1 X Tris Buffered Saline with 0.1% Tween-20 (TBST) for 3 times. The primary antibodies include human VCAM-1 (Santa Cruz, #sc-1504), human ICAM-1 (Santa Cruz, #sc-8439), mouse VCAM1 (R&D, #AF643) and GAPDH (EMD Millipore, #AB2302). Membranes were incubated with appropriate second antibodies at room temperature for 30 min. Odyssey Infrared Imaging System (LI-COR) was used to take images. Densitometric analysis of the blots was analyzed with NIH Image J software.