Xin is a muscle-specific actin binding protein whose role and regulation within skeletal muscle is not well understood. Here we demonstrate that Xin mRNA is robustly upregulated (>16 fold) within 12 hours of skeletal muscle injury and is localized to the muscle satellite cell population. RT-PCR confirmed Xin expression pattern during regeneration, as well as within primary muscle myoblast cultures, but not other known stem cell populations. Immunohistochemical staining of single myofibres demonstrate Xin expression co-localized with the satellite cell marker, Syndecan-4, further supporting the mRNA expression of Xin in satellite cells. In situ hybridization of regenerating muscle 5-7 days post-injury illustrate Xin expression within newly regenerated myofibres. Promoter-reporter assays demonstrate that known myogenic transcription factors (MEF2, MyoD, Myf-5) transactivate Xin promoter constructs supporting the muscle-specific expression of Xin. To determine the role of Xin within muscle precursor cells, proliferation, migration and differentiation analysis using Xin shRNA were undertaken in C2C12 myoblasts. Reducing endogenous Xin expression resulted in a 26% increase (p<0.05) in cell proliferation and a 20% increase (p<0.05) in myoblast migratory capacity. Skeletal muscle myosin heavy chain protein levels were increased (p<0.05) with Xin shRNA administration, however this was not accompanied by changes in myoglobin protein (another marker of differentiation) nor overt morphological differences relative to differentiating control cells. Taken together, the present findings support the hypothesis that Xin is expressed within muscle satellite cells during skeletal muscle regeneration and is involved in the regulation of myoblast function.