Immunohistochemistry.

All reagents for O-GlcNAc and fibronectin immunohistochemistry were obtained from Hisztopatologia (Pecs, Hungary). Slides were deparaffinized in xylene, rehydrated in graded ethanol series, and washed in distilled H₂O. Heat-induced epitope retrieval was performed by boiling the paraffin-embedded tissue sections in citrate buffer (pH 6). Slides were peroxidase blocked, and nonspecific attachments were inhibited with protein solution. Sections were incubated against fibronectin (catalog no. ab2413, Abcam, Cambridge, MA) or O-GlcNAc (shown in Table 1) antibodies followed by peroxidase-labeled anti-rabbit antibody. Fibronectin was visualized with the HISTOLS-Resistant AEC Chromogen/Substrate System counterstained with hematoxylin and eosin and mounted with permanent mounting medium. Evaluation of fibronectin staining was performed similarly to PAS-stained samples.

List of primary antibodies and their use for Western blots

α-SMA; α-smooth muscle actin; CTGF, connective tissue growth factor; EPO, erythropoietin; HIF-1α, hypoxia-inducible factor-1α; OGA, O-GlcNAcase; O-GlcNAc, O-linked N-acetylglucosamine; OGT, O-GlcNAc transferase; PDGF, platelet-derived growth factor; VEGF-A, vascular endothelial growth factor-A; TBS, Tris-buffered saline.