RNA sequencing on FACS-isolated satellite cells 7 days after injury in Tibialis Anterior Muscles

ABSTRACT: The aim of this project is to study satellite cells functionality and get an unprecedented insight into their transcriptional landscape during muscle injury healing process. We aim at revealing their gene expression profiles and describe the continuous variations in their transcriptome after the invalidation of the androgen receptor within their genome. Experimental design: 12 male mice (3 young controls, 3 young mutants, 3 old controls and 3 old mutants) will be injected with cardiotoxin in their Tibialis Anterior Muscles to mimic muscle injury. 7 days after injury, animals will be sacrificed and their tibialis anterior muscles processed in order to FACS-isolate their satellite cells, the stem cells. Stem cells will be immediately sorted in lysis buffer described in the RNeasy Plus Micro Kit protocol. Satellite cells will first be lysed and homogenized in a highly denaturing guanidine-isothiocyanate–containing buffer, which immediately inactivates RNases to ensure isolation of intact RNA. The lysate will then be passed through a gDNA Eliminator spin column. This column, in combination with the optimized high-salt buffer, efficiently removes genomic DNA. Ethanol will be added to the flow-through to provide appropriate binding conditions for RNA, and the sample will then be applied to an RNeasy MinElute spin column, where total RNA binds to the membrane and contaminants will be efficiently washed away. High-quality RNA will then be eluted in as little as 14 µl of water.

ORGANISM(S): Mus musculus

PROVIDER: GSE303612 | GEO | 2026/08/21

REPOSITORIES: GEO