Figure 5.

Uroplakin and tight junction proteins are downregulated by diabetes via NOD-, LRR-, and pyrin domain-containing protein 3 (NLRP3)-dependent mechanisms at the 15-wk (overactive detrusor) time point. Western blots were performed using standard techniques described in methods to determine protein expression of uroplakin subunit UP1b and zona occludens (ZO1) in mucosal layer lysates from all four groups of 15-wk mice. A: representative images showing fluorescent labeled UP1b, ZO1, and GAPDH proteins from all four groups of mice and the respective molecular weights of each protein. B: diabetes downregulated UP1b expression in mice with functional NLRP3. C: in the absence of the NLRP3 gene, diabetic NLRP3^−/− mice demonstrated comparable protein expression as nondiabetic control mice. D: similarly, diabetes downregulated urothelial ZO1 protein. E: without the NLRP3 gene and subsequent NLRP3-mediated inflammation, diabetes did not impact ZO1 protein expression. n = 8 animals per group. *P < 0.05 vs. nondiabetic control mice (Student’s t test).