Coomassie blue stain solution

Mice were sacrificed 7 and 35 days after the rAd injections. Urine, blood and kidney samples were harvested and kept frozen at −80°C. Blood urea nitrogen was assessed using a QuantiChrom Urea Assay kit (BioAssay Systems, DIUR-500) as described previously (Mattison et al., 2012 (link)). The urinary creatinine levels were measured using a colorimetric assay (R&D Systems, KGE005). Albuminuria was measured with a mouse albumin ELISA kit (Bethyl Laboratories, E99-134) and expressed as ratio of the urinary creatinine. In addition, SDS-PAGE (4-12%) was carried out with 5 ml of urine that was then stained with Coomassie blue stain solution (Bio-Rad) to detect changes in high and low molecular weight urinary proteins as described previously (Das et al., 2016 (link)).

Urine, blood and kidney samples were harvested at different time points as described above, and kept frozen at −80°C. BUN was assessed using a QuantiChrom Urea Assay kit (BioAssay Systems, DIUR-500) as described previously (Mattison et al., 2012 (link)). The urinary creatinine levels were measured using a colorimetric assay (R&D Systems, KGE005). Albuminuria was measured with a mouse albumin ELISA kit (Bethyl Laboratories, E99-134) and expressed as a ratio of the urinary creatinine. In addition, SDS-PAGE (4-12%) was carried out with 5 ml urine that was then stained with Coomassie Blue stain solution (Bio-Rad) to detect changes in high- and low-molecular mass urinary proteins as described previously (Das et al., 2016 (link)). For measuring the FGF-2 levels, blood was collected in tubes with EDTA, centrifuged at 4000 g for 5 min and stored immediately at −70°C. FGF-2 secreted by cells transduced with the rAd-FGF-2 vectors was measured with a human FGF basic/FGF-2/Quantikine ELISA Kit (R&D Systems, DFB50), as previously described.