Epithelial to Mesenchymal Transition

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Epithelial to Mesenchymal Transition

Epithelial to Mesenchymal Transition (EMT) describes a mechanism by which cells lose their epithelial characteristics and acquire more migratory mesenchymal properties. This transient and reversible process is classified into three subtypes that are dependent on the biological and functional setting in which it occurs.

Products for EMT Research

EMT Induction and Verification Kits

StemXVivo™ EMT Inducing

Media Supplement

Drives EMT in Cells Resistant to TGF-β

• Rapid - induces EMT in only 5 days

• Versatile - compatible with multiple cell types

• Consistent - defined formulation results in reproducible EMT induction

Human EMT 3-Color Immunohistochemistry Kit

An EMT Research Essential

• Thorough – determines EMT status by protein expression level and subcellular localization

• Efficient – single-step staining using fluorescently-labeled primary antibodies

• Time-Saving – screens for multiple markers simultaneously

Essential Antibodies to Characterize EMT Status

Markers to Monitor EMT

HNF-3β and Cadherin-11 Expression During EMT. The A549 human lung carcinoma cell line was incubated with untreated media (-EMT) or with media containing the StemXVivo EMT Inducing Media Supplement (+EMT; Catalog # CCM017) for 5 days. Cells were stained for the transcription factor HNF-3β/FoxA2 (Catalog # AF2400) and the mesenchymal cell marker Cadherin-11 (Catalog # MAB1790) followed by the NorthernLights™ (NL)557-Conjugated Anti-Goat IgG Secondary Antibody (Catalog #

NL001) and NL493-Conjugated Anti-Mouse IgG Secondary Antibody (Catalog # NL009), respectively. HNF-3β (red) is expressed in untreated A549 cells and decreased in EMT-induced cells. Conversely, Cadherin-11 (green) expression is high in EMT-induced cells and not in untreated cells. The nuclei were counterstained with DAPI (blue).

Upregulated During EMT

Detection of Fibronectin in EMT-Induced Cells. Fibronectin is upregulated in T98G glioblastoma cells induced into EMT (+EMT ) with media containing the StemXVivo EMT Inducing Media Supplement (Catalog # CCM017) compared to untreated cells (–EMT). Fibronectin was detected using the Mouse Anti-Human Fibronectin Monoclonal Antibody (green; Catalog # MAB1918) followed by the NorthernLights™ (NL)493-Conjugated Goat Anti-Mouse Secondary Antibody (Catalog # NL009). The cells were counterstained for E-Cadherin (red) and DAPI (blue). From Tang, Y. et al. (2013) J. Vis. Exp. 78:e50478.

Products to Investigate EMT

EMT Effector Proteins: Highest Purity on the Market

Recombinant Proteins

• Consistent Performance – each lot is tested for consistency to ensure that culture conditions remain the same across experiments • Guaranteed Bioactivity – rigorously tested for high bioactivity using relevant cell culture systems

• World-Class Purity – all proteins meet our industry-leading endotoxin specifications (<0.1 EU/µg)

Functional Assays for EMT: Publication-Ready

Cell Invasion and Migration Assays

• Quantitative – measures cell movement through extracellular matrices

• Flexible – different basement membrane extract (BME) densities and ECM proteins are available

• Published – assays are featured in many peer reviewed journals

Modulators of EMT: Confirmed Bioactivity

Neutralization of TGF-β by Human TGF-β Receptor II Antibody. Addition of Recombinant Human TGF-β1 (rhTGF-β1, Catalog # 240-B) inhibits Recombinant Human IL-4 (rhIL-4)-induced proliferation of TF-1 human erythroleukemic cell line in a concentration dependent manner (green line). The inhibition by rhTGF-β1 was neutralized (orange line) by increasing concentrations of Human TGF-β1 RII Antigen Affinity-Purified Polyclonal Antibody (Catalog # AF-241-NA) with a ND50 of 5–20 µg/mL.

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EMT-Related Small Molecules from Tocris Bioscience

Extracellular Matrix

BMS 536924 - Catalog # 4774

BMS 536924 is a dual inhibitor of the insulin receptor (IR) and insulin-like growth factor-1 receptor (IGF1R) (IC50 values are 73 and 100 nM respectively). In IGF1R

overexpressing MCF10A cells, this compound reverses EMT through the attenuation of Snail mRNA expression and the restoration of E-cadherin protein expression. BMS 536924 also inhibits cell proliferation in multiple tumor types.

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