The collected kidneys were fixed in 10% formalin, dehydrated in graded alcohol series, cleared with xylene, embedded in paraffin, and sectioned at a thickness of 5 μm. Kidney sections were stained with hematoxylin and eosin (H&E). Sirius red staining was used to examine collagen deposition for kidney fibrosis as previously described 16 (link). The proportion of fibrosis (collagen) was quantitated using a Nikon Eclipse E600 microscopy system (Tokyo, Japan).
Standard immunohistochemical (IHC) staining was performed as previously described 17 (link). For the IHC staining, kidney tissue sections were stained with TGF-β, collagen I, collagen IV, laminin, and FN. The sections were developed with a DAB (3,3-diaminobenzidine) developing system (Vector Laboratories, Inc., Burlingame, CA, USA).
Standard immunohistochemical (IHC) staining was performed as previously described 17 (link). For the IHC staining, kidney tissue sections were stained with TGF-β, collagen I, collagen IV, laminin, and FN. The sections were developed with a DAB (3,3-diaminobenzidine) developing system (Vector Laboratories, Inc., Burlingame, CA, USA).
