EHD2 Knockout SK-HEP-1 Polyclonal Cells provide a rigorously validated, CRISPR/Cas9-edited polyclonal knockout cell population designed for precise loss-of-function studies of the EHD2 gene in a human liver sinusoidal endothelial model. Disruption of EHD2 is achieved through CRISPR/Cas9-mediated gene targeting, generating a mixed cellular pool that faithfully reflects the biological heterogeneity of gene knockout effects without selection of single-cell clones. This polyclonal format preserves population-level variation, enabling robust evaluation of EHD2-dependent phenotypes in physiologically relevant contexts.
Hosted in the SK-HEP-1 cell line, which was originally derived from a human hepatic adenocarcinoma but exhibits endothelial characteristics including barrier function, angiogenic capability, and active endocytosis, this model recapitulates key features of liver sinusoidal endothelial cells. SK-HEP-1 cells endogenously express molecular components of the liver endothelium and display functional similarities to primary sinusoidal cells, making them a widely used surrogate for studying hepatic vascular biology and liver-related pathologies.
EHD2 (EH domain-containing protein 2) is a central coordinator of membrane trafficking, particularly caveolae-mediated endocytosis and endocytic recycling of integrins and signaling receptors. Mechanistically, EHD2 oligomerizes around membrane tubules to induce curvature and scission, a process that facilitates caveolar fission and the recycling of cell surface receptors back to the plasma membrane. EHD2 is transcriptionally regulated by SRF and Myocardin and acts downstream of PI3K/Akt signaling. Its function is intimately tied to a network of physical and functional interactors including Caveolin-1, Dynamin-2, Pacsin2/Syndapin, and actin filaments. Within focal adhesion and caveolar pathways, EHD2 is critically positioned upstream of integrin trafficking and caveolae transport, operating in concert with Caveolin-1, PTRF/Cavin-1, Src kinase, FAK, and Rho GTPases to modulate adhesion dynamics and migratory behavior.
In the context of SK-HEP-1 cells, disruption of EHD2 profoundly alters caveolae organization and integrin surface expression, providing a powerful system to dissect the molecular machinery of endothelial endocytosis and its downstream effects on cell adhesion and motility. Because liver sinusoidal endothelial cells serve as a vital interface for nutrient exchange, immune surveillance, and metastatic cell arrest, EHD2 knockout in this model is particularly relevant for investigating the endothelial contribution to cancer metastasis and vascular dysfunction. Researchers can employ this model to study how EHD2 loss impacts signaling receptor recycling, focal adhesion turnover, and transendothelial migration.
This knockout cell population is ideally suited for a broad spectrum of functional assays, including immunofluorescence microscopy to assess caveolae morphology and EHD2/Caveolin-1 colocalization, Western blotting for EHD2 and pathway protein expression, and quantitative migration and invasion assays under standard or stimulated conditions. Additional applications encompass co-immunoprecipitation to confirm protein?Cprotein interactions with Caveolin-1 or Dynamin-2, and flow-cytometric measurement of integrin surface levels to directly gauge recycling efficiency. For further information, please contact Ascent Research.
