Histochemical and Immunohistochemical Analysis of Renal Fibrosis

Histologic and morphologic analysis was performed as previously described [19 (link)]. Histochemical staining was performed with hematoxylin/eosin and sirius red. Immunohistochemical staining was performed using primary antibodies against p-Smad3 (1:500; Santa Cruz Biotechnology, Santa Cruz, CA, USA), α-SMA (1:500; Sigma, St. Louis, MO, USA), PAI-1 (1:500; BD Biosciences, San Jose, CA, USA), and type 1 collagen (1:500; Abcam, Cambridge, UK), followed by horseradish peroxidase-conjugated anti-mouse or anti-rabbit immunoglobulin G secondary antibodies (Dako, Glostrup, Denmark). Quantification of renal fibrosis was measured as previously described [19 (link)].

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Bae K.H., Seo J.B., Jung Y.A., Seo H.Y., Kang S.H., Jeon H.J., Lee J.M., Lee S., Kim J.G., Lee I.K., Jung G.S, & Park K.G. (2017). Lobeglitazone, a Novel Peroxisome Proliferator-Activated Receptor γ Agonist, Attenuates Renal Fibrosis Caused by Unilateral Ureteral Obstruction in Mice. Endocrinology and Metabolism, 32(1), 115-123.

Publication 2017

p-Smad3 α-SMA PAI-1 type 1 collagen horseradish peroxidase-conjugated anti-mouse or anti-rabbit immunoglobulin G secondary antibodies

Anti antibodies Antibodies Eosin Fibrosis Hematoxylin Horseradish peroxidase Immunoglobulin g Mouse Pai 1 Rabbit Renal Smad3 Type 1 collagen

Corresponding Organization : Kyungpook National University Hospital

Other organizations : Keimyung University, Daegu-Gyeongbuk Medical Innovation Foundation

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Variable analysis

independent variables

Histochemical staining with hematoxylin/eosin and sirius red
Immunohistochemical staining with primary antibodies against p-Smad3, α-SMA, PAI-1, and type 1 collagen

dependent variables

Quantification of renal fibrosis

control variables

Not explicitly mentioned

controls

Positive control: Not mentioned
Negative control: Not mentioned

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