For analysis of PPARγ and phospho (p)-NFκB, one retina from each mouse in different groups and treated endothelial cells were homogenized in a modified RIPA buffer (20 mM Tris-HCl [pH 7.4], 2.5 mM ethylenediaminetetraacetic acid, 50 mM NaF, 10 mM Na4P2O7, 1% Triton X-100, 0.1% sodium dodecyl sulfate, 1% sodium deoxycholate, 1 mM phenylmethylsulfonyl fluoride). Homogenates (50 μg protein) were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis using ready precast gel (Bio-Rad, Hercules, CA), transferred to nitrocellulose membrane, and reacted with anti-PPARγ (1:200) and anti–p-NFκB (p65) 1:100 (Santa Cruz Biotechnology, Santa Cruz, CA) followed by horseradish peroxidase-linked secondary antibody and enhanced chemiluminescence (Amersham Pharmacia, San Francisco, CA). Membranes were stripped and reprobed for β-actin or NFκB to demonstrate equal loading, and results were analyzed with the use of densitometry.