Protective Role of Coxsackie-Adenovirus Receptor in the Pathogenesis of Inflammatory Bowel Diseases

Figure 3

CAR overexpression protects human colon cells from TNF-α induced TJ disruption. NCM460 and SW480 cells were transfected with CAR overexpression plasmid and treated with control or TNF-α, and cells were then harvested for qRT-PCR, Western blot, and IF assay detection. (a–d) mRNA levels of cell adhesive marker gene expression ZO-1 (a & b) and Occludin (c & d) upon CAR overexpression detected by qRT-PCR assay. (e) Western-blot examination of protein levels of cell adhesive gene (ZO-1, Occludin, and CAR) upon TNF-α and CAR overexpression in human colon cells lines. β-Actin levels of each sample were used as endogenous loading control. (f–h) Quantitative analysis of CAR, ZO-1, and Occludin protein levels (calibrated with β-actin levels of each sample) in (e). (i) Immunocytochemistry staining detection of ZO-1 protein level after CAR overexpression explored on TNF-α of NCM460 and SW480 cells by IF. , , and ; data represent mean ± s.e.m.

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