Generation and Analysis of Genetically Modified Mice

The generation of p18^-/-, p18^+/-, p18^-/-;Brca1^MGKO (p18^-/-;Brca1^f/f;MMTV-Cre or p18^-/-;Brca1^f/-;MMTV-Cre), p18^-/-;Brca1^+/-, p18^+/-;Brca1^+/-, GATA3^f/f, GATA3^+/-, p18^+/-;GATA3^+/-, and p18^-/-;GATA3^+/- mice has been previously described 7 (link), 15 (link), 20 (link), 48 (link). The Institutional Animal Care and Use Committee at the University of Miami and Shenzhen University approved all animal procedures. Histopathology and immunohistochemistry (IHC) were performed as previously described 7 (link), 15 (link), 20 (link). The primary antibodies used were: BRCA1, E-cadherin (E-Cad), Vimentin (Vim), FRA1 (Cell Signaling), Fibronectin (Fn), TWIST, DNA Methyltransferase 1 (DNMT1) (Abcam), CD29, GATA3 (Santa Cruz). Immunocomplexes were detected using the Vectastain ABC DAB kit according to the manufacturer's instructions (Vector Laboratories). The positive results of IHC were quantified by H-score, as previously described 49 (link), 50 (link).

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Bai F., Zhang L.H., Liu X., Wang C., Zheng C., Sun J., Li M., Zhu W.G, & Pei X.H. (2021). GATA3 functions downstream of BRCA1 to suppress EMT in breast cancer. Theranostics, 11(17), 8218-8233.

Publication 2021

Fibronectin (Fn), DNA Methyltransferase 1 (DNMT1) GATA3 Vectastain ABC DAB kit

Animal Antibodies Brca1 Dna methyltransferase E cadherin Fibronectin Gata3 Immunohistochemistry Institutional animal care and use committee Mice Mmtv Vector Vimentin

Corresponding Organization : University of Miami

Other organizations : Lanzhou University, Henan Cancer Hospital, Zhengzhou University, Second Affiliated Hospital of Xi'an Jiaotong University, University of North Carolina at Greensboro

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Variable analysis

independent variables

P18 genotype (p18-/-, p18+/-, p18-/-)
Brca1 genotype (Brca1MGKO, Brca1+/-)
Gata3 genotype (Gata3f/f, Gata3+/-)

dependent variables

Histopathology
Immunohistochemistry (IHC) for BRCA1, E-cadherin (E-Cad), Vimentin (Vim), FRA1, Fibronectin (Fn), TWIST, DNA Methyltransferase 1 (DNMT1), CD29, GATA3

control variables

Approval of animal procedures by the Institutional Animal Care and Use Committee at the University of Miami and Shenzhen University

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

Annotations

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