Novel Functional Association of Serine Palmitoyltransferase Subunit 1-A Peptide in Sphingolipid Metabolism with Cytochrome P4501A1 Transactivation and Proliferative Capacity of the Human Glioma LN18 Brain Tumor Cell Line
Abstract
:Introduction
Materials and Methods
Chemicals
Cell Culture and Treatment
Antibodies and Staining Reagents
Staining for Immunofluorescence Confocal Microscopy
Immunofluorescence Confocal Imaging of SPT in Cultured Cells
Primer Design, Vectors, RNA Preparation and Quantitative Real Time Reverse Transcriptase PCR (Q-RTPCR)
Functional Engineering and Transfection of SPT1-GFP Vectors
Immunoprecipitation and SDS-polyacrylamide Gel Electrophoresis (SDS-PAGE)
Western Blotting
Cellular Viability/Proliferation Assay
Tandem Mass Spectometry
Results
Endogenous SPT1, Hsp90, SPT1- and AhR-GFP fusion proteins localize to cytoplasmic domain, and 3-MC induces translocation of SPT1 to Focal Adhesion Complex in Glioma LN18 cells
SPT1 is co-precipitated with Hsp90 from lysate of stressed cells and identified in lysate of recombinant cells
Wild type and SPT1 recombinant Glioma cells have similar composition of the long chain sphingoid bases sphinganine (Sa), sphingosine (So) and ceramide (Cer)
The Cyp1A1 gene is differentially induced in wild type and SPT1-GFP recombinant cells following exposure to 3-MC
Discussion
Acknowledgments
References
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Yerokun, T.; Stewart, J. Novel Functional Association of Serine Palmitoyltransferase Subunit 1-A Peptide in Sphingolipid Metabolism with Cytochrome P4501A1 Transactivation and Proliferative Capacity of the Human Glioma LN18 Brain Tumor Cell Line. Int. J. Environ. Res. Public Health 2006, 3, 252-261. https://doi.org/10.3390/ijerph2006030030
Yerokun T, Stewart J. Novel Functional Association of Serine Palmitoyltransferase Subunit 1-A Peptide in Sphingolipid Metabolism with Cytochrome P4501A1 Transactivation and Proliferative Capacity of the Human Glioma LN18 Brain Tumor Cell Line. International Journal of Environmental Research and Public Health. 2006; 3(3):252-261. https://doi.org/10.3390/ijerph2006030030
Chicago/Turabian StyleYerokun, T., and J. Stewart. 2006. "Novel Functional Association of Serine Palmitoyltransferase Subunit 1-A Peptide in Sphingolipid Metabolism with Cytochrome P4501A1 Transactivation and Proliferative Capacity of the Human Glioma LN18 Brain Tumor Cell Line" International Journal of Environmental Research and Public Health 3, no. 3: 252-261. https://doi.org/10.3390/ijerph2006030030