Transforming growth factor beta (TGF-) is a multi-functional cytokine implicated in inflammatory processes, wound healing, and fibrosis. In muscle diseases (i.e. dystrophy and inflammatory myopathy) and in animal models of muscle injury (i.e. produced by cardiotoxin, laceration, and eccentric contractions), increased TGF- was associated with muscle fibrosis and healing. Although TGF- transcript abundance was increased following injury, many studies presume that TGF- protein was also active as evident by increases in collagen transcript abundance. The purpose was to determine whether TGF- protein is present and active 48 hours following injury. Using female rats, muscle strains were produced by stretching (50 stretches) the plantar flexor muscles. Forty-eight hours following injury, the medial gastrocnemius was removed and compartmentalized into 5 equal segments. Damaged myofibers with intracellular Concanavalin A staining were counted. The percent of damaged myofibers was significantly greater in the distal most segment. TGF- was assessed using immunohistochemistry, reverse transcriptase polymerase chain reaction (RT-PCR) and immunoblot analysis. Immunohistochemistry revealed the presence of TGF-₁ in areas of myofiber injury, whereas TGF-₂ was not detected. Increases in TGF-₁ and TGF-₂ transcript abundance following strain injury were documented by RT-PCR analysis. Increases in TGF-₁ and TGF-₂ precursor abundance were observed following strain injury using immunoblot analysis but there was no change in active TGF- abundance. Although there was no correlation between the amount of cellular injury and TGF- transcript and protein abundance, elevated levels of TGF-₁ and TGF-₂ precursor proteins were present in strain-injured skeletal muscles 48 hours after injury.