Immunohistochemical Analysis of Tissue Samples

Tissue samples were fixed and paraffinized as described^{25 (link)}. The 4 μm thick sections were prepared from paraffin-embedded tissues and immunostained with anti-CD45 (X16/99, Novocastra, Leica biosystems, Newcastle, UK, Cat# NCL-L-LCA), anti-Notch3 (Santa Cruz Biotechnology, Dallas, TX, USA, Cat#5539) or anti-CHOP (Cell Signalling, Danvers, MA, USA, Cat#2895) antibodies. After washes, secondary biotinylated antibodies were applied. Binding of antibodies was detected with the Mouse to Mouse HRP (DAB) Staining System (Scytek Laboratories, Inc., Logan, UT, USA) according to the manufacturer’s protocol. The analysis was conducted blindly.

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Giuli M.V., Diluvio G., Giuliani E., Franciosa G., Di Magno L., Pignataro M.G., Tottone L., Nicoletti C., Besharat Z.M., Peruzzi G., Pelullo M., Palermo R., Canettieri G., Talora C., d’Amati G., Bellavia D., Screpanti I, & Checquolo S. (2020). Notch3 contributes to T-cell leukemia growth via regulation of the unfolded protein response. Oncogenesis, 9(10), 93.

Publication 2020

anti-Notch3 anti-CHOP

Antibodies Chop Embedded paraffin Mouse Notch3 Tissues

Corresponding Organization :

Other organizations : Sapienza University of Rome, Sylvester Comprehensive Cancer Center, University of Miami, Novo Nordisk Foundation, Italian Institute of Technology, Rutgers, The State University of New Jersey

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Variable analysis

independent variables

Tissue samples were fixed and paraffinized

dependent variables

Immunostaining with anti-CD45, anti-Notch3, or anti-CHOP antibodies

control variables

Tissue samples were prepared from paraffin-embedded tissues
Analysis was conducted blindly

controls

Positive control: Not specified
Negative control: Not specified

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