Protein Extraction and Western Blot Analysis

Proteins were extracted from cell lines using RIPA Buffer (Thermo) according to the manufacturer's protocol. Protein extraction and western blot analysis with chemiluminescent detection were as described [50 (link)]. The following antibodies and dilution factors were used: FOXM1 rabbit polyclonal antibody (13147-1-AP, 1:800, Proteintech), cyclin B1 (CCNB1) rabbit polyclonal antibody (4138P, 1:800, Cell Signaling), anti-rabbit vimentin (5741, 1:100 Cell Signaling), anti-rabbit E-cadherin (3195, 1:200, Cell Signaling), mouse GAPDH monoclonal antibody (MA5-15738, 1:2000, Sigma), anti-rabbit IgG conjugated to horse radish peroxidase (7074S, 1:2000, Cell Signaling), and anti-mouse IgG (7076S, 1:2,000, Cell Signaling).

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Tan X., Fu Y., Chen L., Lee W., Lai Y., Rezaei K., Tabbara S., Latham P., Teal C.B., Man Y.G., Siegel R.S., Brem R.F, & Fu S.W. (2015). miR-671-5p inhibits epithelial-to-mesenchymal transition by downregulating FOXM1 expression in breast cancer. Oncotarget, 7(1), 293-307.

Publication 2015

RIPA Buffer anti-rabbit vimentin (5741 anti-rabbit E-cadherin anti-mouse IgG (7076S

Anti igg Antibodies Antibody Buffer Cadherin Cell lines Cyclin b1 Dilution Gapdh Horse radish peroxidase Monoclonal antibody Mouse Protein Rabbit Ripa Vimentin Western blot analysis

Corresponding Organization :

Other organizations : George Washington University, Bon Secours Health System

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Protein expression levels of FOXM1, cyclin B1 (CCNB1), vimentin, E-cadherin, and GAPDH

control variables

Cell lines used for protein extraction
RIPA Buffer used for protein extraction
Antibody dilution factors

controls

Positive control: None explicitly mentioned
Negative control: None explicitly mentioned

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