CCT2 Expression in Carcinoma Tissue Microarrays

Tissue microarrays (TMAs) used in this study were obtained from US Biomax, Inc. Their catalog numbers are as follows: CO484a (colonic carcinoma), PR803b and PR631 (prostate carcinoma), BC03118 (hepatocellular carcinoma), LC726b, LC802a, LC802c, and BC041115a (lung carcinomas) and BN501 (normal tissue array). Each TMA contained varied numbers of patient tissue cores as well as normal tissue corresponding to the specific cancer type being analyzed. Please refer to Supplementary Tables 1, 2 and 5 for the number of samples per cancer type. Information about the tissue type, TNM, score, tumor grade, and stage were provided with the samples. TMAs were stained for CCT2 using anti-CCTβ antibody (LS-B4861; LifeSpan Biosciences). TMA LC802c was stained in parallel for CCT2 and Stat3 (anti-Stat3 antibody ab32500; Abcam). Primary antibodies were diluted 1:100 in Antibody Diluent (Leica). Staining of tissue arrays was performed by a Bond-Max Immunostainer (Leica), with an epitope retrieval buffer of EDTA pH 9.0 (Leica). Polymer Refine Detection reagents (Leica) were used, which include a hematoxylin counterstain. Image acquisition and scoring of staining was performed by a surgical pathologist as previously published [31 (link)].

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Carr A.C., Khaled A.S., Bassiouni R., Flores O., Nierenberg D., Bhatti H., Vishnubhotla P., Manuel J.P., Santra S, & Khaled A.R. (2017). Targeting chaperonin containing TCP1 (CCT) as a molecular therapeutic for small cell lung cancer. Oncotarget, 8(66), 110273-110288.

Publication 2017

LS-B4861 ab32500 Antibody Diluent Bond-Max Immunostainer Polymer Refine Detection reagents

Anti antibody Antibodies Antibody Buffer Cancer Colonic carcinoma Edta Epitope Hematoxylin Hepatocellular carcinoma Lung carcinomas Microarrays Pathologist Patient Polymer Prostate carcinoma Staining tissue Stat3 Surgical Tissue Tissue type Tumor

Corresponding Organization : University of Central Florida

Other organizations : Orlando VA Medical Center, Cedars-Sinai Medical Center, Pittsburg State University

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Variable analysis

independent variables

Tissue type (colonic carcinoma, prostate carcinoma, hepatocellular carcinoma, lung carcinomas, normal tissue)

dependent variables

CCT2 expression (as measured by immunohistochemical staining)
Stat3 expression (as measured by immunohistochemical staining)

control variables

Antibody dilution (1:100)
Epitope retrieval buffer (EDTA pH 9.0)
Staining method (Bond-Max Immunostainer)
Detection reagents (Polymer Refine Detection)
Hematoxylin counterstain

controls

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

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