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Fig. 2 | Molecular Medicine

Fig. 2

From: Lysozyme Enhances Renal Excretion of Advanced Glycation Endproducts In Vivo and Suppresses Adverse AGE-mediated Cellular Effects In Vitro: A Potential AGE Sequestration Therapy for Diabetic Nephropathy?

Fig. 2

Lysozyme enhances macrophage binding, internalization and degradation of AGE formed in vitro or in human sera. (A)

Confluent cultures of murine macrophage-like T1B-183 cells (0.5 ×106/well) were incubated with either 125I-AGE-BSA, 125I-LZ or 125I-BSA (10–25 µg/well), in the presence or absence of 100-fold excess of the indicated unlabeled ligand, for 2 hrs at 4°C. (B) LZ was immobilized on a NC membrane (10 µg/dot) and probed with 125I-MG-BSA (4 × 106 cpm, or 5 µg/ml) in the presence or absence of 50-fold excess of unlabeled AGE-BSA, MG-BSA, CML-BSA or BSA for 1 hr. Radiolabeled ligand binding was visualized by exposure to XAR film (Kodak) at −80°C (upper), and quantitated by phosphorimage analysis of three identical ligand dot blots, expressed as M ± SD (lower). (C) T1B-183 cells, plated as in (A) were incubated with either 125I-labeled AGE-BSA (100 µg/well) or 125I-human AGE-enriched serum (150 µg/well) with or without LZ (80 µg/ml) for 4 hrs at 37°C. (D) Media from T1B-183 cell cultures treated as in (C) were collected and after TCA-precipitation, soluble cpm were determined. For (A) (C) and (D) panels, data are expressed as the M ± SD cpm of five independent experiments, each performed in triplicate. *p < 0.05, **p < 0.01 vs. LZ.

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