Quantification of Glomerular Remodeling

Mouse kidney tissues were fixed in 4% paraformaldehyde and embedded in paraffin. Periodic acid Schiff (PAS) staining was performed on 3 μm sections. All glomeruli (between 50–100 glomeruli) in a single transverse section for each mouse were examined under a microscope at 400× total magnification and scored individually in a blinded manner and then averaged. Semiquantitative scores (0–4+) were assigned based on blinded readings. Mesangial matrix expansion or sclerosis scoring was performed as we described previously [18 (link), 22 (link), 25 (link)–27 (link)]. Images were captured with an Olympus BX51 microscope and DP71 digital camera using cellSens Standard 1.12 image software. Images were obtained with 100× (oil) objective with a total magnification of 1000×. Immunohistochemistry for macrophages was performed using rat anti-mouse Mac-2 antibody (clone M3/38; Cedarlane, Burlington, NC) on paraffin sections as described previously [18 (link), 25 (link)–27 (link)].

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Awad A.S., You H., Gao T., Gvritishvili A., Cooper T.K, & Tombran-Tink J. (2015). Delayed Treatment with a Small Pigment Epithelium Derived Factor (PEDF) Peptide Prevents the Progression of Diabetic Renal Injury. PLoS ONE, 10(7), e0133777.

Publication 2015

rat anti-mouse Mac-2 antibody (clone M3/38;

Anti antibody Camera Clone Embedded paraffin Glomeruli Immunohistochemistry Kidney Macrophages Mesangial Microscope Mouse Paraffin Paraformaldehyde Periodic acid Sclerosis Tissues

Corresponding Organization : Pennsylvania State University

Top 5 similar protocols

Variable analysis

independent variables

Mesangial matrix expansion or sclerosis scoring

dependent variables

Semiquantitative scores (0–4+) of mesangial matrix expansion or sclerosis
Macrophage immunohistochemistry

control variables

Mouse kidney tissues were fixed in 4% paraformaldehyde and embedded in paraffin
Periodic acid Schiff (PAS) staining was performed on 3 μm sections
All glomeruli (between 50–100 glomeruli) in a single transverse section for each mouse were examined under a microscope at 400× total magnification and scored individually in a blinded manner and then averaged
Immunohistochemistry for macrophages was performed using rat anti-mouse Mac-2 antibody (clone M3/38; Cedarlane, Burlington, NC) on paraffin sections

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