Total RNA was isolated from frozen gastrocnemius muscle using the RNeasy Mini Kit (Qiagen, Hilden, Germany) according to the manufacturer's instructions. RNA concentration and purity were assessed using a NanoPhotometer, and RNA integrity was evaluated using an Agilent 2100 Bioanalyzer. A total of 800 ng RNA with an RNA integrity number of at least 7.0 was used for library preparation. Poly(A)-positive RNA was isolated using the NEBNext Poly(A) mRNA Magnetic Isolation Module. Directional sequencing libraries were prepared using the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina according to the manufacturer's instructions. Library quality and fragment size were assessed using an Agilent 2100 Bioanalyzer. Libraries were sequenced on an Illumina NovaSeq 6000 in paired-end mode with a read length of 2 x 61 bp, yielding approximately 43.0-53.8 million reads per sample.