RNA-sequencing of soleus muscle from Mustn1 heterozygous KO-first mice and wildtype littermates
To uncover novel genes and molecular pathways important for skeletal muscle remodeling and regeneration, we used a mouse hindlimb unloading and reloading protocol. We then performed analysis of global gene expression by RNA-sequencing (RNA-seq), looking at the transition from muscle unloading to reloading (GSE237099). This study focuses on the microprotein Mustn1 (Musculoskeletal embryonic nuclear protein 1, also known as Mustang), which consists of 82 amino acids. We performed RNA-sequencing of soleus muscle from 8-week-old mice carrying one allele (heterozygous) of Mustn1 tm1a (KO-first) and wildtype littermates. Overall design: RNA-sequencing analysis of soleus muscles from Mustn1 KO-First Heterozygous (HET) mice and wild-type (WT) littermate controls at 8 weeks of age.  Total RNA was extracted using TRI reagent (Sigma-Aldrich ref. T9424) according to the manufacturer's instructions. RNA was further purified using a Direct-zol RNA Miniprep kit (Zymo Research) including on-column DNAse I treatment. RNA quality was assessed using a 2100 Bioanalyzer system (Agilent Technologies) with an RNA integrity number (RIN) greater than 8 as a threshold. Samples were purified by poly-A enrichment before library preparation using Illumina TruSeq Stranded mRNA and sequenced on Illumina HiSeq2500 (single reads, 50 bp, 30M reads per sample).