Immunohistochemical staining was performed on paraffin‐embedded tissue, as described previously 36 . Immunostaining for α‐SMA and CD3 was accomplished with the EnVision System (Dako, Glostrup, Denmark). Other markers were identified with the ABC system (Vector Laboratories). The sections were all visualized using 3,3′‐diaminobenzidine (Sigma–Aldrich). CD3‐, CD68‐ and EGFP‐positive cells and the positive areas for α‐SMA, collagen type I, and collagen type III staining were assessed as the average of five randomly selected fields (×200) for each rat with ImageJ software (NIH).
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Immunohistochemistry Protocol for Tissue Analysis
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Corresponding Organization : Hiroshima University
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Variable analysis
independent variables
- Immunohistochemical staining
- CD3-positive cells
- CD68-positive cells
- EGFP-positive cells
- Positive areas for α-SMA
- Positive areas for collagen type I
- Positive areas for collagen type III
- Paraffin-embedded tissue
- EnVision System (Dako, Glostrup, Denmark)
- ABC system (Vector Laboratories)
- 3,3'-diaminobenzidine (Sigma–Aldrich)
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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