Analysis of O -GlcNAcylated cytoskeletal proteins by using the in situ PLA assay. Localization of O-GlcNAcylated actin (A, B), O-GlcNAcylated α-actinin 4 (C, D), O-GlcNAcylated α-tubulin (E, F) and O-GlcNAcylated myosin (G, H) in the glomerulus (1) and tubule (2) of normal (A, C, E, G) and diabetic (B, D, F, H) rats. Arrows in D indicate podocytes. I-L) Relative number of signals per cell. Ten different glomeruli and tubules were obtained from each sample. Signals were analyzed by Blob-Finder software. Values represent means ± SEM from 3 different rats. *P < 0.05 vs. control Wistar rat (W).