Immunohistochemical Analysis of FTSEC Tumors

Immunohistochemistry was performed on consecutive 4-µm-thick sections of formalin fixed and paraffin embedded FTSEC mouse tumors. Antigen retrieval and staining for gamma H2AX (clone and manufacturer?) was performed as previously described in ref. ^{57 (link)}. Staining for PAX8 (Roche, clone MRZ-50) and Ki-67 (Roche, clone 30-9) was performed on an automated BenchMark ULTRA staining module (Ventana) using ULTRA CC1 cell conditioning for antigen retrieval and the OptiView DAB Detection Kit (Ventana). All slides were analyzed in a blinded fashion by a gynecological pathologist.

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Knarr M., Avelar R.A., Sekhar S.C., Kwiatkowski L.J., Dziubinski M.L., McAnulty J., Skala S., Avril S., Drapkin R, & DiFeo A. (2020). miR-181a initiates and perpetuates oncogenic transformation through the regulation of innate immune signaling. Nature Communications, 11, 3231.

Publication 2020

BenchMark ULTRA staining module OptiView DAB Detection Kit

Antigen Cell Clone Formalin Gamma Immunohistochemistry Mouse Paraffin Pathologist Pax8 Tumors

Corresponding Organization :

Other organizations : Case Western Reserve University, University of Michigan–Ann Arbor, University of Pennsylvania

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Variable analysis

independent variables

Tissue samples: Formalin fixed and paraffin embedded FTSEC mouse tumors

dependent variables

Expression of gamma H2AX
Expression of PAX8
Expression of Ki-67

control variables

Consecutive 4-µm-thick sections of the tissue samples
Antigen retrieval and staining protocols for gamma H2AX, PAX8, and Ki-67
Automated BenchMark ULTRA staining module (Ventana) for PAX8 and Ki-67 staining
Analysis of slides in a blinded fashion by a gynecological pathologist

controls

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

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