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Detzner, Johanna ; Krojnewski, Elisabeth; Pohlentz, Gottfried; Steil, Daniel ; Humpf, Hans-Ulrich ; Mellmann, Alexander ; Karch, Helge; Müthing, Johannes: Shiga Toxin (Stx)-Binding Glycosphingolipids of Primary Human Renal Cortical Epithelial Cells (pHRCEpiCs) and Stx-Mediated Cytotoxicity
Inhalt
Results
Identification of Stx1a- and Stx2a-Binding Globo-Series Glycosphingolipids of pHRCEpiCs
Structural Characterization of Sphingolipids from the Neutral GSL Fraction of pHRCEpiCs
Lipid Composition of DRM and Non-DRM Fractions Obtained from pHRCEpiCs
Mass Spectrometric Specification of the Stx Receptor GSLs Gb3Cer and Gb4Cer in DRM and Non-DRM Fractions Obtained from pHRCEpiCs
Mass Spectrometric Specification of the Phospholipids and SM in DRM and Non-DRM Fractions Obtained from pHRCEpiCs
Stx1a- and Stx2a-Mediated Cellular Damage of pHRCEpiCs
Discussion
Conclusions
Materials and Methods
Cultivation and Propagation of pHRCEpiCs
Stx Cytotoxicity Assay
Harvest of DRM and Non-DRM Fractions from Sucrose Density Gradients
Isolation and Purification of GSLs from Total Cells
Obtaining Phospholipid and GSL Preparations from DRM and Non-DRM Fractions
Stx1a, Stx2a, Antibodies, and Lipid References
Thin-Layer Chromatography, Lipid Staining, and Overlay Detection
Mass Spectrometry of GSLs
References