Kidney samples were placed in 4% paraformaldehyde, followed by being segmented at a thickness of 4 μm from paraffin-embedded tissues. Then hematoxylin and eosin (H&E) were carried out to assess pathological renal injury, and Masson’s trichrome staining and Sirius Red staining to observe collagen deposition. Renal tubule injury severity was determined with five grades (0–4): 0, no obviously visible injury; 1, injury <25%; 2, injury 25–50%; 3, injury 50–75%; and 4, injury >75% [40 (link)–42 (link)].
As for immunohistochemical staining, antibodies, included anti-RCAN1 (1:200, D6694, Sigma-Aldrich), anti-YY1 (1:200, ab109228, Abcam), anti-E-cad (1:100, #40860, SAB), anti-TNF-α (1:200, ab6671, Abcam), anti-α-SMA (1:100, 14395-1-AP, Proteintech), were employed to incubate kidney section, followed by an appropriate secondary antibody. The stained samples were observed using an Ortho microscope (OLYMPUS, Tokyo, Japan).
As for immunohistochemical staining, antibodies, included anti-RCAN1 (1:200, D6694, Sigma-Aldrich), anti-YY1 (1:200, ab109228, Abcam), anti-E-cad (1:100, #40860, SAB), anti-TNF-α (1:200, ab6671, Abcam), anti-α-SMA (1:100, 14395-1-AP, Proteintech), were employed to incubate kidney section, followed by an appropriate secondary antibody. The stained samples were observed using an Ortho microscope (OLYMPUS, Tokyo, Japan).
Free full text: Click here
