Figure 5

WNV-induced CAM expression in the brains of WT and db / db mice. (A) qRT-PCR was conducted on RNA extracted from mock- and WNV-infected brains from WT and db/db mice at indicated time points to determine the fold-change in E-selectin, ICAM-1, and VCAM-1 gene expression. Changes in the levels of each gene were first normalized to the β-actin gene, and then the fold-change in WNV-infected brains was calculated in comparison to corresponding mock-infected brains. Data represents the mean ± SEM, representing two independent experiments (n = 7 per group). *P <0.05. (B) The protein levels of E-selectin, ICAM-1 and VCAM-1 were analyzed using Western blot analysis. Total brain lysates were separated by SDS-PAGE, transferred onto nitrocellulose membranes and immunoblotted with antibodies specific to E-selectin, ICAM-1, and VCAM-1. Equal loading was confirmed by re-probing with anti-β-actin antibody and the bands were detected using the Li-Cor Odyssey infrared method. (C) Quantitative analysis of Western blots results. Changes in the levels of each gene was first normalized to the β-actin gene and then the percentage increase in WNV-infected brains was calculated in comparison to corresponding mock-infected brains. Data represents the mean ± SEM, representing two independent experiments (n = 7 per group). *P <0.05. ICAM-1, intercellular cell adhesion molecule 1; VCAM-1, vascular cell adhesion molecule 1; SEM, standard error of mean.