Recombinant mouse klotho protein

Thirty-eight-week-old C57BL/6 male mice were supplied from Shanghai Sipper-BK Laboratory Animal Co. (Shanghai, China). All procedures were approved by the Animal Care and Use Committee of our hospital. All animals were housed with a 12-h light/dark cycle and free access to water and standard commercial rat chow. Mice were randomly designated into three groups (n = 6 per group): sham, model, and Klotho treatment. Renal fibrosis was induced by UUO, as described in our previous study (Liu et al., 2015 (link)). All experimental mice underwent the UUO procedure except for mice in the sham group (whereby the ureter was not obstructed). Recombinant mouse Klotho protein (R&D systems) was administered to mice in the Klotho treatment group by intraperitoneal injection at a dosage of 0.02 mg/kg every other day starting immediately after UUO operation (Liu et al., 2015 (link)). Mice in sham and model groups were injected with physiological saline. Body weight was monitored and the dosage of agents was calculated based on body weight. On day 14 after UUO operation, blood samples were collected and obstructed kidneys were rapidly removed from mice (under anesthetization) and washed with saline. Obstructed kidneys were processed for histopathological examination, immunofluorescence, immunohistochemistry, and western blotting analysis.