A siRNA Mediated Screen During C2C12 Myogenesis.
Methods Mol Biol
; 1889: 229-243, 2019.
Article
em En
| MEDLINE
| ID: mdl-30367417
ABSTRACT
Myogenesis is a multistep process taking place during pre- and postnatal stages for muscle formation, growth, and regeneration. It is a highly regulated process involving many molecular factors which act during myoblast proliferation and differentiation. To provide new insights into the molecular mechanisms and interactions behind the regulation of these different steps, RNA interference is an efficient methodology to implement. We developed a high-throughput siRNA screen in C2C12 murine myoblast cells for identification of genes relevant to signaling pathways controlling muscle growth. The proposed protocol is based on (1) the analyses of a maximum number of cells/myotubes to detect and quantify both clear and subtle phenotypes during proliferation/fusion cells and (2) the use of two cellular fluorescent markers, DAPI and myosin, decorating nuclei and myotubes respectively. Four phenotypic criteria were quantitatively assessed cellular density, myotubes quantity, fusion index, and size and morphology of myotubes.
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Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Desenvolvimento Muscular
/
Mioblastos
/
RNA Interferente Pequeno
Idioma:
En
Ano de publicação:
2019
Tipo de documento:
Article
País de afiliação:
França