Adult 8-week-old female CD1 mice were purchased from the Charles River Laboratory (Wilmington, MA, RRID: IMSR_CRL:22). Mice were anesthetized with 1%–5% isoflurane gas with continuous inhalation. The second and fourth digits of both hind limbs were amputated at the P3 distal level as previously described and regenerating digits were collected at days 14, 21, 28, and 42 for analysis (Sammarco et al., 2015 (link); Busse et al., 2019 (link); Hoffseth et al., 2021a (link); Hoffseth et al., 2021b (link); Tower et al., 2022 (link)). The third digit was used as an unamputated control. Oxaloacetate (OAA) (Sigma, 07753) was dissolved in saline (PBS) and pH-adjusted with NaOH to 7.0 to allow the OAA solution to drift toward alkaline levels as previously described (Tower et al., 2022 (link)), with the exception that mice were treated daily with 1 g/kg OAA in 200 µL via i.p. injection. The vehicle-treated group received saline. Mice were dosed from day 10 until day 28. Dichloroacetate (DCA) (Alfa, B21897) was dissolved in PBS. Mice were treated daily with 0.05 g/kg DCA (Liang et al., 2020 (link)) in 100 µL via i.p. injection. The vehicle treated group received PBS.
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