We followed the wound induced hair neogenesis protocol as published in Ito et al. Nature 2007. We waited 5-weeks after injury to assess for nascent hair follicles. To detect newly developing hair follicles in the wound, we incubated the skin in 20 mM EDTA in PBS at 37° C overnight. We peeled off the epidermis and fixed the dermis in acetone at 4° C overnight, rinsed it with PBS, and then incubated with NBT/BCIP (Roche) for 30 min at 37° C. We stopped the reaction with 20 mM EDTA in PBS.