Western Blotting Analysis of Key Markers

SDS-PAGE and western analyses were completed as previously described^{89 (link),90 (link)}. Briefly, normalized protein samples were loaded onto either 8% or 10% SDS-PAGE gels (as appropriate) and western blotting was completed with the following antibodies from Cell Signaling Technology (Danvers, MA, USA): (1) EVI1 rabbit monoclonal (#2593, 1:500), (2) Myc rabbit monoclonal (#13987, 1:500), (3) total Ras rabbit monoclonal (#3339, 1:1000), (4) β-catenin rabbit polyclonal (#9587, 1:1000), (5) BMI1 rabbit monoclonal (#6964, 1:1000), (6) LC3B rabbit polyclonal (#2775, 1:1000), and (7) Pan-Actin rabbit polyclonal (#4968, 1:500). SV40 LTAg mouse monoclonal (#554149, 1:1000) and p62 mouse monoclonal (#610832, 1:1000) antibodies were obtained from BD Biosciences (San Jose, CA, USA). FANCD2 mouse monoclonal (#sc-20022, 1:1000), Cyclin D1 rabbit polyclonal (#sc-718, 1:1000), and FoxJ1 mouse monoclonal (#sc-365216, 1:500) were obtained from Santa Cruz Biotechnology.

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Rockfield S., Kee Y, & Nanjundan M. (2019). Chronic iron exposure and c-Myc/H-ras-mediated transformation in fallopian tube cells alter the expression of EVI1, amplified at 3q26.2 in ovarian cancer. Oncogenesis, 8(9), 46.

Publication 2019

LC3B rabbit polyclonal Pan-Actin rabbit polyclonal SV40 LTAg mouse monoclonal Cyclin D1 rabbit polyclonal

Actin Antibodies Bmi1 Cyclin d1 Evi1 Fancd2 Gels Mouse Protein Rabbit Sds page Sv40 β catenin

Corresponding Organization :

Other organizations : University of South Florida

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Variable analysis

independent variables

SDS-PAGE gel percentage (8% or 10%)

dependent variables

Protein expression levels of: EVI1, Myc, total Ras, β-catenin, BMI1, LC3B, Pan-Actin, SV40 LTAg, p62, FANCD2, Cyclin D1, FoxJ1

control variables

Normalized protein samples

controls

Positive controls: Not explicitly mentioned
Negative controls: Not explicitly mentioned

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