Your browser doesn't support javascript.
loading
Flow cytometry protocol for GLUT4-myc detection on cell surfaces.
Zanni-Ruiz, Emilia; Mayorga, Luis Segundo; Pavarotti, Martin Alejandro.
Afiliação
  • Zanni-Ruiz E; Laboratorio de Transporte Intracelular, Instituto de Histología y Embriología de Mendoza Dr. Mario H Burgos, Mendoza, Argentina.
  • Mayorga LS; Consejo Nacional de Investigaciones Científicas y Técnicas, Buenos Aires, Argentina.
  • Pavarotti MA; Universidad Nacional de Cuyo, Mendoza, Argentina.
Biosci Rep ; 44(4)2024 Apr 24.
Article em En | MEDLINE | ID: mdl-38533799
ABSTRACT
Insulin and muscle contraction trigger GLUT4 translocation to the plasma membrane, which increases glucose uptake by muscle cells. Insulin resistance and Type 2 diabetes are the result of impaired GLUT4 translocation. Quantifying GLUT4 translocation is essential for comprehending the intricacies of both physiological and pathophysiological processes involved in glucose metabolism. The most commonly used methods for measuring GLUT4 translocation are the ELISA-type assay and the immunofluorescence assay. While some reports suggest that flow cytometry could be useful in quantifying GLUT4 translocation, this technique is not frequently used. Much of our current understanding of the regulation of GLUT4 has been based on experiments using the rat myoblast cell line (L6 cell) which expresses GLUT4 with a myc epitope on the exofacial loop. In the present study, we use the L6-GLUT4myc cell line to develop a flow cytometry-based approach to detect GLUT4 translocation. Flow cytometry offers the advantages of both immunofluorescence and ELISA-based assays. It allows easy identification of separate cell populations in the sample, similar to immunofluorescence, while providing results based on a population-level analysis of multiple individual cells, like an ELISA-based assay. Our results demonstrate a 0.6-fold increase with insulin stimulation compared with basal conditions. Finally, flow cytometry consistently yielded results across different experiments and exhibited sensitivity under the tested conditions.
Assuntos
Palavras-chave

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Músculo Esquelético / Diabetes Mellitus Tipo 2 Limite: Animals Idioma: En Revista: Biosci Rep Ano de publicação: 2024 Tipo de documento: Article País de afiliação: Argentina País de publicação: Reino Unido

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Músculo Esquelético / Diabetes Mellitus Tipo 2 Limite: Animals Idioma: En Revista: Biosci Rep Ano de publicação: 2024 Tipo de documento: Article País de afiliação: Argentina País de publicação: Reino Unido