*4.6. Western Blotting*

Western blotting analysis was performed as previously described [28]. Briefly, protein from the obstructed kidney tissue or cells was extracted in a radioimmunoprecipitation assay buffer containing a protease inhibitor cocktail (cOmplete-Mini, Roche, Indianapolis, IN, USA). The protein concentration was determined by a Bradford assay (Bio-Rad Laboratories, Montreal, Quebec, Canada), separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and, subsequently, transferred to polyvinylidene fluoride membranes. The membranes were then probed with primary antibodies against fibronectin (1:1000, Cat#15613-1-AP, Proteintech Group, Chicago, IL, USA), α-SMA (1:5000, Cat#14395-1-AP, Proteintech Group), iNOS (1:1000, Cat#ab3523, Abcam), Arg1 (1:1000, Cat#93668S, Cell Signaling Technology, Boston, MA, USA), CLEC7A (1:500, Cat#TA322197, OriGene Technologies), SLAM (1:1000, Cat#ab156288, Abcam), TNF-α (1:1000, Cat#ab66579, Abcam) and β-actin (1:5000, Cat#60008, Proteintech Group) at 4 ◦C overnight. Afterwards, the membranes were incubated with horseradish peroxidase-conjugated secondary antibodies (Jackson ImmunoResearch, West Grove, PA, USA) at room temperature for 1.5 h, and the signals were developed using a West Femto Chemiluminescent Substrate kit (Thermo Fisher Scientific, Hudson, NH, USA). Bands were visualized and quantified using a ChemiDoc-It Imaging system (UVP, Cambridge, UK). Data were normalized to the β-actin expression.
