Colonic Mucosa Histopathology and Immunohistochemistry

For all experimental groups, formalin-fixed and paraffin-embedded colonic mucosa sections were stained with hematoxylin and eosin for conventional histopathological analysis. Immunohistochemical staining for PCNA and β-catenin were performed using the labeled streptavidin-biotin method (LSAB kit; Dako, Glostrup, Denmark), as previously described [14 (link),24 (link),25 (link)]. The primary antibodies for β-catenin and PCNA were obtained from BD Transduction Laboratories (No. 610154; San Jose, CA, USA) and Santa Cruz Biotechnology (sc-7907; Santa Cruz, CA, USA), respectively. PCNA-positive cells in the colonic mucosa were counted and expressed as a percentage of the total number of normal crypt cells. The PCNA labeling index (%) was determined from the assessment of a minimum of 200 crypt cells in each mouse [29 (link)].

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Fukuta K., Shirakami Y., Maruta A., Obara K., Iritani S., Nakamura N., Kochi T., Kubota M., Sakai H., Tanaka T, & Shimizu M. (2017). Preventive Effects of Pentoxifylline on the Development of Colonic Premalignant Lesions in Obese and Diabetic Mice. International Journal of Molecular Sciences, 18(2), 413.

Publication 2017

LSAB kit sc-7907

Antibodies Biotin Cells Colonic Crypt Eosin Formalin Mouse Mucosa Paraffin Pcna Streptavidin β catenin

Corresponding Organization : Gifu University

Other organizations : Gifu Municipal Hospital

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

PCNA-positive cells in the colonic mucosa (expressed as a percentage of the total number of normal crypt cells)
PCNA labeling index (%)

control variables

Formalin-fixed and paraffin-embedded colonic mucosa sections
Hematoxylin and eosin staining for conventional histopathological analysis
Immunohistochemical staining for PCNA and β-catenin using the labeled streptavidin-biotin method (LSAB kit; Dako, Glostrup, Denmark)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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