FIG. 11.

CXCR4 mediates AOPP-induced podocyte injury in glomerular miniorgan culture. (A) Representative micrograph shows glomerular miniorgan culture in vitro. Glomeruli were isolated from rat kidneys and cultivated in suspension. (B–D) Western blotting shows that AOPPs induced CXCR4 upregulation and inhibited nephrin expression in cultured glomeruli. Rat glomeruli were treated with AOPPs (0–50 μg/ml) for 24 h. Quantitative analysis show CXCR4 (C) and nephrin (D) expression in different groups as indicated. *p < 0.05 versus controls (n = 3). (E–G) Western blotting shows that AOPPs induced CXCR4 upregulation and suppressed nephrin expression, which were abolished by AMD3100 in cultured glomeruli. Rat glomeruli were pretreated with AMD3100 (5 μg/ml) for 1 h and then treated with AOPPs (50 μg/ml) for 24 h. Quantitative analyses show CXCR4 (F) and nephrin (G) expression in three groups. *p < 0.05 versus controls (n = 3). ^†p < 0.05 versus AOPPs alone (n = 3). (H) Immunofluorescence staining shows that AOPPs inhibited expression of nephrin and WT1, which was abolished by AMD3100 in glomerular culture. Arrows indicate positive staining. Scale bar, 40 μm. All full unedited gels for Figure 11 are presented in Supplementary Figure S15.