The transforming growth factor beta (TGFβ)/activin signaling pathway has a number of documented roles during wound healing and is becoming increasingly appreciated as a vital component of multi-tissue regeneration. The leopard gecko (Eublepharis macularius) is able to spontaneously, and repeatedly, regenerate its tail following tail loss. We thus examined the expression and localization of several key components of the TGFβ/activin signaling pathway during tail regeneration of the leopard gecko. We observed a marked increase in phosphorylated-Smad2 expression among regenerating tissues corresponding to the location of the regenerate blastema. Interestingly, we observe that during early regeneration there appears to be an absence of TGFβ family member TGFβ1 and instead a strong upregulation of activin-βA. We also observe the expression of EMT transcription factors Snail1 and Snail2 in blastemal tissue. These observations combined with other data provide strong support for the importance of unique and non-overlapping expression patterns of different TGFβ ligands during multi-tissue regeneration