*4.5. Muscle RNAseq Analysis*

Previously, we performed RNAseq analysis on gastrocnemius muscle mRNA in 12-month-old CKD mice versus age-appropriate sham control mice [23]. Detailed procedures for mRNA extraction, purification, and subsequent construction of cDNA libraries as well as analysis of gene expression were published. We then performed ingenuity pathway analysis enrichment tests for those differentially expressed muscle genes in CKD mice versus control mice, focusing on pathways related to energy metabolism, skeletal and muscle system development and function, and organismal injury and abnormalities. We identified the top 12 differentially expressed muscle genes in CKD versus control mice. In this study, we performed qPCR analysis for those top 12 differentially expressed gastrocnemius muscle genes in the different experimental groups.
