Deficiency of IKKα in Macrophages Mitigates Fibrosis Progression in the Kidney after Renal Ischemia-Reperfusion Injury

Figure 7

Primary TECs treated with CoCl₂ to imitate IRI in vitro and then cocultured with WT MΦs or KO MΦs. Then, the fibrosis markers and Wnt/β-catenin pathway were investigated. (a) Expression of α-SMA by immunohistochemistry in control TECs (top) and IRI TECs (bottom) cocultured with PBS (left), WT MΦs (middle), and KO MΦs (right). Representative images of α-SMA staining show distribution in the cytoplasm of TECs that underwent fibrosis. Magnification, ×100. (b) Immunohistochemistry images were measured by ImageJ, and data are presented as the . versus control and coculture with KO MΦs; ^# versus TECs without IRI; . (c) Protein levels of α-SMA, vimentin, and collagen I as fibrosis markers were measured by Western blotting. (d) Protein levels of β-catenin, active β-catenin, and Snai1 as signals of the Wnt/β-catenin pathway were measured by Western blotting. (e) Quantification of Western blot analyses for the proteins mentioned above; the data showed that all fibrosis markers were increased significantly accompanied by Wnt/β-catenin pathway activation in IRI TECs cocultured with KO MΦs compared with the WT MΦ coculture group and control group, while the expression of the proteins showed no significant difference in TECs without IRI. Data are presented as the . versus control or coculture with KO MΦs; ^# versus TECs without IRI; , one-way ANOVA followed by the Student-Newman-Keuls test.

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