Histopathological and Immunohistochemical Analysis of Renal Tissue

Renal tissue samples were collected from mice and were fixed in 4% paraformaldehyde, embedded in paraffin and 4 µm thick serial sections were obtained. Then, the sections were deparaffinized, hydrated, and stained with hematoxylin and eosin (H & E) in order to assess histopathological kidney injury and Sirus-red and Masson’s trichrome staining for observing collagen deposition. On the basis of the procedure described in previous research, immunohistochemical staining of RCAN1.4 was performed [48 (link)]. The following primary antibody of rabbit anti-RCAN1 (1:200 dilution, pH 8.0, Sigma-Aldrich, USA) was used. The stained tissues were viewed by Ortho microscope (OLYMPUS, Tokyo, Japan).