proteome profiler human protease array kit  (R&D Systems)

Journal: Biomedicines

Article Title: Extracellular Vesicles Derived from Human Liver Stem Cells Counteract Chronic Kidney Disease Development and Cardiac Dysfunction in Remnant Kidney Murine Model: The Possible Involvement of Proteases

doi: 10.3390/biomedicines12071517

Figure Lengend Snippet: Protease expression profile in EVs. ( A ) Representative image of the results of Proteome Profiler TM Human Protease Array, comparing the expression of 35 human proteases among HLSC and HLSC-EV protein lysates. The top 13 proteases expressed in cells and EVs are listed in the table on the right. ( B ) Histogram representing the average signal (pixel density of the spots) of the 13 proteases expressed in cell protein lysate (blue bars) and in EV lysate (orange bars). ( C ) Representative image of the results of Western blot analyses of 5 proteases expressed in the protein lysate of cells and EVs purified by ultracentrifugation (UC-EVs) and EVs purified by floating (FL-EVs). ( D ) Histological quantification of fibrosis in SHAM mice, PNx mice treated with vehicle alone, and PNx mice injected with dose 1 of EVs pre-treated or not with blocking antibody (n = 6/group). Results are shown as mean ± SD. ( E ) Representative images of renal sections of PNx mice treated with vehicle alone or with dose 1 of EVs pre-treated or not with specific blocking antibody stained with Masson’s trichrome. The blue stain represents collagen fibers. Original magnification: 400×. Bar scale: 50 μm.

Article Snippet: To characterize the expression profile of 35 human proteases in HLSCs and HLSC-EVs, the Proteome Profiler TM Human Protease Array Kit (ARY021B, R&D Systems, Minneapolis, MN, USA) was used, following the manufacturer’s instructions.

Techniques: Expressing, Western Blot, Purification, Injection, Blocking Assay, Staining