Expression of EGR1 gene and location of EGR1 protein in differentiation of bovine skeletal muscle-derived satellite cells / 中国应用生理学杂志
Chinese Journal of Applied Physiology
; (6): 5-8, 2019.
Article
в Zh
| WPRIM
| ID: wpr-776569
Ответственная библиотека:
WPRO
ABSTRACT
OBJECTIVE@#To investigate the expression of EGR1 gene and the localization of EGR1 protein in bovine skeletal muscle-derived satellite cells (MDSCs), as well as to investigate the mechanism that EGR1 protein enters the nucleus.@*METHODS@#Bovine MDSCs were cultured in differentiation medium for 1 day, 3 days and 5 days, respectively, and each group was triplicate. The expression of EGR1 gene and the localization of EGR1 protein were studied at different differentiation period in MDSCs by qRT-PC and Western blot. Moreover, the changes on the expression of endogenous EGR1 gene and EGR1 proteins were explored by CRISPRi, site-directed mutagenesis and laser confocal method.@*RESULTS@#The results from the qRT-PCR and Western blot showed that the expressions of EGR1 gene on transcription level and translation level were significantly higher in differentiated cells than those in undifferentiated cells. The highest expression was found on the third day after the differentiation, and then began to decline. Immunofluorescence assays showed that EGR1 proteins were preferentially expressed in differentiated MDSCs, and increased along with the increase of number of myotubes. Confocal observation revealed that some EGR1 proteins were transferred into the nucleus in the differentiation of cells, however, the EGR1 proteins would not be detected in the differentiated MDSCs nuclei if a site directed mutagenesis (serine) on EGR1 protein occurred.@*CONCLUSION@#During the differentiation of bovine skeletal muscle satellite cells, the transcriptional level of EGR1 gene is increased, and some EGR1 proteins are transferred into the nucleus. The serine phosphorylation at position 533 of the C terminal of EGR1 protein is necessary for the nucleus transfer.
Полный текст:
1
База данных:
WPRIM
Основная тема:
Cell Differentiation
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Cell Nucleus
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Cells, Cultured
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Muscle Fibers, Skeletal
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Satellite Cells, Skeletal Muscle
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Early Growth Response Protein 1
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Genetics
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Metabolism
Пределы темы:
Animals
Язык:
Zh
Журнал:
Chinese Journal of Applied Physiology
Год:
2019
Тип:
Article